Modified Cry34 proteins

ABSTRACT

This invention provides modified, insecticidal Cry34 proteins with enhanced properties as compared to wild-type Cry34 proteins. The modifications to these proteins were based in part on an analysis of the three-dimensional (3D) structure of this protein and other proteins in the Cry34 class. The subject invention also includes polynucleotides that encode these modified proteins, and transgenic plants that produce these modified proteins. This invention further provides methods of controlling plant pests, including rootworms, with these modified proteins. The modified proteins of the subject invention include chimeric toxins involving exchanged segments, domains, and motifs as discussed herein. The subject invention also provides methods of modifying Cry34 proteins.

CROSS-REFERENCE TO A RELATED APPLICATION

The present application is a divisional of U.S. Ser. No. 10/956,725,filed Oct. 1, 2004, which claims benefit to Provisional Application Ser.No. 60/508,567, filed Oct. 3, 2003, which are hereby incorporated byreference herein in their entirety, including any figures, tables,nucleic acid sequences, amino acid sequences, or drawings.

BACKGROUND OF THE INVENTION

Coleopterans are a significant group of agricultural pests that causeextensive damage to crops each year. Examples of coleopteran pestsinclude corn rootworm and alfalfa weevils. Additional notable examplesinclude Colorado potato beetle, boll weevil, and Japanese beetle.

Insecticidal crystal proteins from some strains of Bacillusthuringiensis (B.t.) are well-known in the art. See, e.g., Höfte et al.,Microbial Reviews, Vol. 53, No. 2, pp. 242-255 (1989). These proteinsare typically produced by the bacteria as approximately 130 kDaprotoxins that are then cleaved by proteases in the insect midgut, afteringestion by the insect, to yield a roughly 60 kDa core toxin. Theseproteins are known as crystal proteins because distinct crystallineinclusions can be observed with spores in some, strains of B.t. Thesecrystalline inclusions are often composed of several distinct proteins.

A new insecticidal protein system was discovered in Bacillusthuringiensis as disclosed in WO 97/40162. This system comprises twoproteins—one of approximately 15 kDa and the other of about 45 kDa. Seealso U.S. Pat. Nos. 6,083,499 and 6,127,180. These proteins have nowbeen assigned to their own classes, and accordingly received the Crydesignations of Cry34 and Cry35, respectively. See Crickmore et al.website (biols.susx.ac.uk/home/Neil_Crickmore/Bt/). Many other relatedproteins of this type of system have now been disclosed. See e.g. U.S.Pat. No. 6,372,480; WO 01/14417; and WO 00/66742. Plant-optimized genesthat encode such proteins, wherein the genes are engineered to usecodons for optimized expression in plants, have also been disclosed. Seee.g. U.S. Pat. No. 6,218,188.

Details of the three-dimensional structure of these proteins have not,heretofore, been disclosed. With information regarding thethree-dimensional structures of these proteins, it would be possible torationally design modifications to the natural, bacterial proteins toimprove various desirable characteristics of these proteins. Having andanalyzing the 3D structure of a protein can be highly advantageous forfocusing or restricting directed evolution and improvement programs.

However, obtaining purified crystals of B.t. insect toxins has been adifficult process (although some examples do exist; see e.g. WO 98/23641and WO 99/31248). It has been difficult to obtain purified crystals ofadequate quality. For example, there has been a tendency for theseproteins to form aggregates that are not suitable for refinement of thestructure to high resolution. In addition, B.t. has been an inferiorprotein producer for the level and quality of protein required for X-raycrystallography and related biochemical purposes. Frequent proteasecontamination has also been an associated obstacle. Still further,native B.t. strains typically produce crystals having a mixture ofproteins; thus, there have been some issues with isolating and purifyingsingle protein types from such mixtures (to the degree required forsophisticated analysis).

BRIEF SUMMARY OF THE INVENTION

This invention provides modified, insecticidal Cry34 proteins withenhanced properties as compared to wild-type Cry34 proteins. Themodifications to these proteins as discussed below were based in part onan analysis of the three-dimensional (3D) structure of the ˜15 kDa 149B1protein and other proteins in the Cry34 class. The subject inventionalso includes polynucleotides that encode these modified proteins, andtransgenic plants that produce these modified proteins. This inventionfurther provides methods of controlling plant pests, includingrootworms, with these modified proteins.

The modified proteins of the subject invention include chimeric toxinsinvolving exchanged segments, domains, and motifs as discussed herein.

The subject invention also provides methods of modifying Cry34 proteins.However, the modifications described herein can be applied to other(structurally similar) proteins and peptides as well.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B illustrate the basic three-dimensional structure oftypical Cry34 proteins.

FIGS. 2A and 2B provide an illustration of the three-dimensionalstructure of typical Cry34 proteins, with some surface-exposed residuesindicated.

FIGS. 3A and 3B illustrate surface-exposed residues with an additionallevel of detail, and can also be used to show the two main domains ofCry34 molecules.

FIG. 4 shows a multiple sequence alignment of various Cry34 proteins.

BRIEF DESCRIPTION OF THE SEQUENCES

SEQ ID NO:1 is the amino acid sequence of the wild-type 149B1 ˜15 kDa(Cry 34Ab1) protein.

SEQ ID NO:2 is an example of a dibasic residue truncation ormodification to improve activity according to the subject invention.

SEQ ID NO:3 is an example of a dibasic residue truncation ormodification to improve activity according to the subject invention.

SEQ ID NO:4 is an example of a dibasic residue truncation ormodification to improve activity according to the subject invention.

SEQ ID NO:5 is an example of a dibasic residue truncation ormodification to improve activity according to the subject invention.

SEQ ID NO:6 is an example of using a run of glycines to reduce effect ofMet1 hydrophobicity.

SEQ ID NO:7 is an example of using a run of glycines to reduce effect ofMet1 hydrophobicity.

SEQ ID NO:8 is an example of using a run of glycines to reduce effect ofMet1 hydrophobicity.

SEQ ID NO:9 is the Cry34 protein designated PS201HH2.

SEQ ID NO:10 is the Cry34 protein designated PS201L3.

SEQ ID NO:11 is the Cry34 protein designated PS185GG.

SEQ ID NO:12 is the Cry34 protein designated PS69Q.

SEQ ID NO:13 is the Cry34 protein designated PS80JJ1.

SEQ ID NO:14 is the Cry34 protein designated KR1369.

SEQ ID NO:15 is the Cry34 protein designated PS167H2.

SEQ ID NO:16 is the Cry34 protein designated PS158×10.

SEQ ID NO:17 is the Cry34 protein designated PS149B1.

BRIEF DESCRIPTION OF THE APPENDICES

Appendix 1 provides the atomic coordinates for the 149B1 Cry34 protein.

Appendix 2 is a spreadsheet that includes accessibility informationregarding the amino acid residues of Cry34Ab1.

DETAILED DESCRIPTION

This invention provides modified, insecticidal Cry34 proteins withenhanced properties as compared to wild-type Cry34 proteins. Themodifications to these proteins as discussed below were based in part onanalysis of the three-dimensional (3D) structure of the ˜15 kDa 149B1protein and other proteins in the Cry34 class, together with otheranalytic approaches. The subject invention also includes polynucleotidesthat encode these modified proteins, and transgenic plants that producethese modified proteins, and seeds and other plant materials (such aspollen and germplasm) produced by such plants. This invention furtherprovides methods of controlling plant pests, including rootworms, byusing these modified proteins.

As referred to herein, Cry34-M proteins are any proteins modified orproduced synthetically (that differ from wild-type Cry34 proteins)according to the methods disclosed and/or suggested herein.

Synthetic proteins of the subject invention include Cry34-M proteinswith increased stability in plants and/or increased activity againstinsects.

Some synthetic proteins of the subject invention have one or more aminoacid substitutions that improve binding, protease resistance (in plants,for example) and/or susceptibility (in insect guts, for example),hydrophobicity/hydrophilicity, charge distribution, and likecharacteristics of the synthetic proteins as compared to wild-type Cry34proteins.

Some synthetic proteins of the subject invention are the result ofmodifying one or more amino acid residues of a given wild-type Cry34protein (a Cry34A protein, for example) to make the resulting syntheticsequence more or less like that of a different wild-type Cry34 protein(a Cry34B protein, for example). This approach was based in part onsubstituting residues based on sequence diversity in homologous proteintoxins together with analyzing the corresponding crystal structure.

The modified proteins of the subject invention include chimeric toxinsinvolving exchanged domains and motifs as discussed herein.

Further proteins of the subject invention are obtainable by focusedsequence shuffling or site saturation mutagenesis, wherein saidshuffling is directed, as described herein, to certain regions orsegments of Cry34 proteins.

Still further, proteins of the subject invention include those that wereobtained in part by using computational molecular evolution based inpart on structural data. That is, while sequence alignments/comparisonsof various Cry34 proteins can provide some clues as to differencesbetween given proteins in this class, sequence alignments alone are notable to convey similar structural motifs that might be shared by variousproteins, including Cry34-class proteins.

The subject invention includes methods of modifying at least one aminoacid residue of a Cry34 protein, including the step of consulting athree-dimensional model of a Cry34 protein.

Atomic coordinates for the 149B1 Cry34 protein are provided in Appendix1.

Basic Structure of Cry34 Proteins

FIGS. 1A and 1B illustrate the basic structure of the Cry34 proteins.FIGS. 2A and 2B illustrate a further level, with some surface-exposedresidues indicated in the illustration. FIGS. 3A and 3B provide a stillfurther detailed illustration. FIGS. 3A and 3B basically suggest thatthe molecule is in two modules: a front subdomain (approximatelyresidues 1-67) and a back subdomain (residues ˜67-end). As discussedbelow in Example 6, this also suggests that the subdomain boundary wouldbe a good place to make chimerics.

Before discussing the various structural features and overall structureof the Cry34 molecules, it should be noted that “˜” used before a rangeof numbers (e.g., ˜1-9) signifies that this is an approximate range ofresidues (unless otherwise specified). Thus, ˜1-9 means the same as ˜1-9unless otherwise indicated. Some examples of overlapping segmentdefinitions can be found herein.

The overall structure of Cry34 molecules can be summarized as follows.Some residues omitted at the ends (residues ˜1-2 and ˜120-123) areassumed to be a part of the amino acid chain in the crystals, but theyare too variable in position to be fixed in the model.

Strand # amino acid residues location/orientation or loop of segment onFIG. 1 1 ~4-13 bottom-top Loop ~14-18  top 2a, 2b ~19-21; 25-27top-bottom Loop ~28, 29  bottom 3 ~30-32  bottom-top coil 33-41bottom-top Loop ~39-41  top 4 ~42-50  top-bottom 4-5 loop ~51-56  bottom5 ~57-65  bottom-top Loop ~66-68  top 6 ~69-77  top-bottom 6-7 loop~78-83  bottom 7 ~84-90  bottom-top Loop ~91-92  top 8 ~93-101top-bottom Loop ~102-103  bottom 9 ~104-115  bottom-top coil 116-119 topcoil 120-123 top - not observed in crystal

Residues ˜1-9 form a beta strand running (N terminus to C terminaldirection) from the bottom to top of the Cry34 molecule as illustratedin FIG. 1. A loop occurs at residues ˜14-18 (at the top of the moleculeas illustrated in FIG. 1) followed by strand 2 (residues ˜19-21; 25-27),consisting of two short beta strands, which runs downward as illustratedin FIG. 1.

This is followed by another loop at residues ˜28-29 (bottom FIG. 1).Residues ˜30-41 form a segment (running back up the molecule of FIG. 1)consisting of a short beta strand (30-32) and a less structured segment(33-41).

Segment ˜42-50 is a beta strand running (N->C) back down the molecule asshown in FIG. 1.

The large loop, at the bottom of the molecule of FIG. 1, extending fromresidues ˜50-57 is very interesting. This is discussed in more detailbelow.

The ˜58-68 segment runs back to top of molecule (as illustrated inFIG. 1) where there is a loop at residues ˜66-70.

The ˜70-78 segment (strand 6) runs back down to the ˜78-81 loop. The˜81-91 segment (strand 7) transitions into a ˜91-95 loop at the top ofthe molecule of FIG. 1.

Strands 6-7 are involved with the formation of a center pore, asdiscussed in more detail below. As such, the inward-facing residues inthese strands are preferably not modified. Similarly, the ˜76-80 loop ispreferably not modified.

The ˜95-102 segment travels back down the molecule to a “bottom” loop atresidues ˜102-106.

The segment of residues ˜106-114 travels back up the molecule and endsat the carboxy terminus at ˜123, after the protruding tail at the topleft of the molecule of FIG. 1 (after residue ˜114).

Possible Mechanisms of Action of Cry34 Proteins

The Cry35 protein is known to act with the Cry34 (˜15 kDa) protein. The3D structure of the Cry35 protein is discussed in more detail in U.S.Ser. No. 60/508,637 entitled, “Modified Cry35 Proteins.” Without beinglimited by any one theory, the Cry34 protein could bind to a multimericassociation of assembled Cry35 proteins via a cross-subunit bindingsite. This would explain the inability of Cry34/35 to form associationsin vitro in initial observations. (Thus, it appears unlikely that amembrane-bound Cry35 monomer associates with the membrane and then withthe 14 kDa as a binding partner.) It would be consistent with otherknown, similar protein models if the Cry35 multimer associates with thecellular membrane and embeds using a beta-hairpin-based membraneinteraction domain. Upon multimerization, this could form abeta-barrel-like assembly of the Cry35 subunits—usually seven. (The betahairpin of Cry35 is from residues ˜238-262, centered at 254 and 255, andis structurally similar to other proposed hairpins for other knownproteins. Although sequence similarity with those proteins is weak,there is structural similarity, which also suggests that the bottomloops, especially ˜78-83, embed in the membrane.) The multimer wouldthen facilitate entry of the 15 kDa protein, which could have a cellulartarget via binding, or could form pores on its own (i.e. beta-barreltype via a loop of residues ˜28-˜55).

It appears that the Cry34 protein could insert into insect cellmembranes. One manner in which this could occur, based on variousmolecular and energetic analyses discussed herein, is via “16-39unfolding.” “Hinging out” of the segment comprising strands 2-3 wouldexpose the hydrophobic core of this protein to the membrane surface.Strands 2-3 can thus be thought of as the bar of a hand grenade, whichsprings out when it is not depressed. While not being limited by asingle theory regarding an exact mechanism of action, one possibility isthat multiple ˜15 kDa proteins could associate and form a channel inthis manner. As illustrated, and in this model, the C-terminal tailsticks straight up and could bind the ˜45 kDa (Cry35).

A second model involves residues 27-53 (strands 3-4). This model isinteresting because the 3∃ strands are long enough to span the membrane.Although the remainder of the molecule in this conformation does notappear to be very stable, the 30-50 segment could fold onto the othersheet.

Yet another model involves residues ˜15-56 (strands 2-3 and 3-4). Thisis a more variable portion of the sequence in the Cry34 family,especially residues ˜27-53 (strands 3-4). One option is to modify aresidue in this segment to turn it into an amphipathic ∀-helix. Thestretch from residues ˜42-57 has a distinct ∀/∃ hydrophobic moment. Itis also possible to observe some alpha helical amphipathic character onhelical wheel slots of the 30-53/55 stretch.

In any case, the loops between strands 2 and 3 (residues 28-29) and 4and 5 (residues 51-56) are key hinges.

For residues that are identified herein as being ideal for substitution,conservative changes can be made as defined below in Example 8. However,in some cases, nonconservative changes would be preferred. The efficacyof such changes can be initially analyzed using computer modeling suchas Voigt, C. A., Mayo, S. L., Arnold, F. H., and Wang, Z. G.,“Computationally focusing the directed evolution of proteins,” J. CellBiochem. (2001), Suppl. 37:58-63; and Voigt, C. A., Mayo, S. L., Arnold,F. H., and Wang, Z. G., “Computational method to reduce the search spacefor directed protein evolution,” Proc. Natl. Acad. Sci. U.S.A. (Mar. 27,2001), 98(7):3778-83. Techniques for producing and confirming theactivity of proteins modified accordingly are well-known in the art.

It should be understood that while the specific residue numbers referredto herein relate primarily to the exemplified 149B1 protein, the subjectdisclosure shows that all Cry34 proteins have similar structures tothose exemplified herein. Thus, as one skilled in the art would know,with the benefit of this disclosure, corresponding residues and segmentsare now identifiable in the other Cry34 proteins. Thus, the specificexamples for the 149B1 protein can be applied to the other proteins inthe Cry34 family. The exact numbering of the residues might not strictlycorrespond to the 149B1 protein, but the corresponding residues arereadily identifiable in light of the subject disclosure. See, e.g., FIG.4.

Unless indicated otherwise herein, all known Cry34 wild-type proteinsappear to have the same basic structure, although there are someimportant differences in their amino acid residues at certain positions.The sequences of various Cry34 proteins and genes are described invarious patent and other references as indicated below (such sequencescan be used according to some embodiments of the subject invention): Forexample, the following protein sequences can be used according to thesubject invention:

Cry designation Source isolate GENBANK Acc. No. 34Aa1 PS80JJ1 AAG5034134Aa2 EG5899 AAK64560 34Ab1 PS149B1 AAG41671 34Ac1 PS167H2 AAG5011834Ac2 EG9444 AAK64562 34Ba1 EG4851 AAK64566

35Aa1, 35Ab1, and 35Ac1 are also disclosed in WO 01/14417 as follows.

SEQ ID NO: Source isolate IN WO 01/14417 PS80JJ1 32 PS167H2 36 PS149B141

There are many additional Cry34 sequences disclosed in WO 01/14417 thatcan be used according to the subject invention. For example:

SEQ ID NO: Source isolate IN WO 01/14417 PS131W2 52 PS158T3 56 PS158X1060 PS185FF 62 PS185GG 66 PS185L12 70 PS185W3 72 PS186FF 74 PS187F3 76PS187L14 84 PS187Y2 88 PS204G4 100 PS204I11 102 PS204J7 104 PS236B6 106PS242K10 108 PS246P42 112 PS69Q 114 KB54 118 KR1209 120 KR1369 122 KR589124 PS201L3 134 PS187G1 138 PS201HH2 142 KR1369 146 PS137A 150 PS201V2152 PS207C3 154Several other source isolates are also disclosed in WO 01/14417. The PSdesignation of the source isolate can be dropped for ease of referencewhen referring to a protein obtainable from that isolate. Variouspolynucleotides that encode these proteins are also known in the art anddisclosed in various references cited herein.

All patents, patent applications, provisional applications, andpublications referred to or cited herein are incorporated by referencein their entirety to the extent they are not inconsistent with theexplicit teachings of this specification.

Following are examples that illustrate procedures for practicing theinvention. These examples should not be construed as limiting. Allpercentages are by weight and all solvent mixture proportions are byvolume unless otherwise noted.

Example 1 Modification of Surface-Exposed Residues

The following table lists exposed residues and the degree to which theyare exposed:

Residue # Degree of exposure 3 some exposure 22 some exposure 23outward-facing side chain 25 some exposure 27 outward-facing side chain28 outward-facing side chain 29 outward facing side chain 30 outwardfacing side chain 33 more exposed 34 more exposed 39 more exposed 44more exposed 51 outward-facing side chain 52 outward-facing side chain53 outward-facing side chain/some exposure 54 outward-facing sidechain/more exposed 66 more exposed 67 more exposed 80 outward-facingside chain 81 outward-facing side chain 82 outward-facing side chain 83more exposed/some exposure 91 more exposed 92 some exposure inoutward-facing side chain 102 outward-facing side chain 105 someexposure 116 outward-facing side chain 117 some exposure 118 someexposure 119 some exposure

In general, these residues (especially those that are “more exposed” and“outward facing”) are preferred for modification and would have littleimpact on the overall structure of the molecule. That is, if function isaffected, the modification of function would be due most likely to thealteration of the (exposed) side chain, as opposed to a propagatedstructure distortion elsewhere.

Example 2 Modification of Exposed Loops and “Charged Girdle”Modifications to Improve Solubility

A “charged girdle” can be identified above the hydrophobic bottom loops.Histidines in the “girdle” can be changed to R or K to improvesolubility. Likewise, the “T” at position 60 can be changed to H, K, orR (or E). Following are other examples of changes that can be made toimprove the solubility of the molecule:

H7R, H16R, H88R, H107R, and N51H.

Alternatively, an H7Y modification can be for improved stability. Thus,preferred Cry35-M proteins have histidine residues modified to R or K.

Another possibility is V47 (an outward facing hydrophobic residue) toH,K,R, or more generally to I, M, L, T, A, K, H, or R.

Preferably, for all of the modifications suggested herein (in thisExample and elsewhere throughout), single changes would be made first,and then multiple changes would be made—combining the singlemodifications that result in equal or better activity.

Example 3 Scanning-Based Changes

Appendix 2 provides data that was analyzed to determine residues thatwould be good to change, based on similarity value (less is better),accessibility (more is better), outward facing side chain (more isbetter), and B-factor (how well fixed the residues are in the crystalstructure (more is better). Using the table of Appendix 1, accessibleresidues with high B-factors (i.e. >30 in last column) were initiallyidentified, then accessible residues with scores of 1 & 2, thenoutfacing with high B-factor, then outfacing score 2, and then thosewith an outfacing score of 1. Substitutions can follow those found inthe different families, prioritized by profile similarity (substitutionscolumn), etc.

Example 4 Nearest Neighbor Analysis

A nearest neighbor analysis of the first 55 residues was conducted.H-bonds from residue 56 and higher were also identified that: 1) connectto 16-36, and 2) are between sheets (68-89 connecting to 94-110). Thisanalysis indicated that the residues past 56 were more interconnected byH-bonds than the earlier segment. Residues S34 and N51 do not havenon-adjacent neighbors and should be highly substitutable. That is, S34and N51 have minimal contacts aside from adjacent residues and should behighly substitutable. Changing S34 is preferred.

Example 5 Analysis of Force-Field Energies

An analysis was conducted of force-field energies and threading energyrankings. Higher energies relative to the electron density data couldindicate stress on the protein that could aid unfolding. Segments 20-24,33-36 and 43-44 are potentially stressed and worthy of modification. Byusing this design, one can obtain a molecule that behaves normally but“unfolds” easier when desired.

Example 6 Construction of Chimerics

Chimerics can be constructed according to the subject invention toassess functionality, preferably residues 66/67 as the crossover point,and preferably using the 201 L3 14 kDa gene (as the 201L3 14 kDa is muchless active, this can show which of the segmental sequence differencesdisclosed herein are responsible, as well as where large numbers ofchanges are tolerated). Additional chimerics with, for example, 80JJ1and 158×10 can also be constructed to assess activity and stabilityeffects.

Chimerics of the subject invention can also be truncated as explainedbelow in Example 7. These combinations can be constructed to assess theeffects of different (or omitted or truncated) homolog C-termini,including the effects of charge and polarity changes. Thus, preferredchimerics are of the 1-66/67-end type.

Example 7 Terminal Deletions to Produce Truncated Cry 34 Proteins

5′ and 3′ deletions can be performed to make N- and C-terminallytruncated proteins. The essential minimum coding segment can bedetermined in this manner.

In addition, as plant-produced protein is minus Met1, improvement ofactivity could be obtained from N-terminal truncation or modificationswith, e.g., dibasic residues (e.g. MKKSAREVH (SEQ ID NO.:2) . . . ,MKKAREVH (SEQ ID NO.:3) . . . , or MGGGSAR (SEQ ID NO.:4) . . . , MGGGAR(SEQ ID NO.:5). . . ) to enhance cleavage to get S2 or A3 at theN-terminus. Alternatively, the subject invention includes the use of arun of glycines to reduce effect of Men hydrophobicity. A3 or R4 appearto be critical, so the truncations would encode MAREVH (SEQ ID NO.:6) .. . , MREVH (SEQ ID NO.:7), MEVH (SEQ ID NO.:8) . . . , etc. untilactivity is drastically affected.

3′ deletions can also be constructed, and the criticality of the tailcan be assessed. Residues prior to T114 appear to be critical, soterminal truncations of whole segments could advantageously be made,rather than processively from the 3′ end one at a time. These techniquescan also be used to determine the functionality of family variability inthe C-termini (T114 on), as much of this may be totally dispensible.

Alternatively, the C-terminus can be retained, but cross-over chimericscan be constructed in this region to improve activity. In the Cry34family, there are several sequence variants in this region with only anArg (R118) totally conserved. One example of this type of variant wouldconsist of the 149B1 sequence through T114, then 80JJ1 sequence (forexample) could be used at the terminus. Various combinations of thistype could be constructed using any of the Cry34 family members.

Truncations that exhibit improved activity or other functionality orcharacteristics can also be used with further approaches to modificationand improvement as discussed above and elsewhere herein (and viceversa).

Example 8 Analyzing Multiple Sequence Alignments

Thus, according to the guidance provided herein, one can align andcompare the sequences of any or all known Cry34 homologues. Onealignment of some Cry34 alleles is shown in FIG. 5. Chemical propertiesof the residues can be compared in such an alignment, and then relatedto the 3D structure. This type of combined analysis can now beconducted.

Another method of the subject invention is to, for example, introduceany one or more or all possible changes observed (from such alignments)in the other related Cry34 proteins to the Cry 34Ab1 protein, forexample, if these changes are in regions of the protein that wouldtolerate change, based on an analysis of the 3D structure of theproteins as disclosed herein. Conversely, the subject invention includesmaking the 201L3 protein more like another Cry 34 protein, such as the149B1 Cry 34 protein, if these changes are in regions of the proteinthat would tolerate change, based on an analysis of the 3D structure ofthe proteins as disclosed herein. The 201L3 binary toxins are the mostdivergent, by sequence, and are also less active than the 149B1 binarytoxins; however, the 201L3 14 kDa protein, for example, is moresusceptible to protease processing than is the 149B1 protein.

Unless otherwise indicated, sequences were aligned using ClustalWdefault parameters at the ClustalW WWW Service at the EuropeanBioinformatics Institute website (ebi.ac.uk/clustalw). Various sequenceanalysis software is available for displaying various alignments,including the free Genedoc package available at(psc.edu/biomed/genedoc/). Multiple sequence alignments can be analyzedusing two Genedoc functions:

Conservation mode produces a display that emphasizes the degree ofconservation in each column in the alignment. Positions with 60, 80 or100% identity, for example, can be shaded in different grayscale tones.Residue similarity scoring can be enabled, such that residue similaritygroups (Blossum 62) are given arbitrary numbers on the consensus line.

Chemical properties highlights sequence residues that share a definedset of properties. In this analysis default shading can be used tohighlight the following groups by color:

negatively positively charged charged amide alcohol aliphatic aromaticsmall sulfur other D, E H, K, R N, Q S, T L, I, V F, Y, W A, G M, C PResidue substitutions can be identified by scanning the length of thesequence alignment. Thus, one can align the sequences of various Cry34proteins and look for “outlying” amino acids (residues that aredifferent, i.e. of a different chemical class, as compared to others ata corresponding position).

Again, the 149B1 and 201L3 Cry34 proteins are good reference points, inpart because the 149B1 Cry34/Cry35 combination is one of the most activebinary toxin combinations (wild-type) known to date. On the other hand,the 201L3 Cry34/Cry35 combination is one of the most active binary toxincombinations (wild-type) known to date.

Using the atomic coordinates and guidance provided herein, one canconduct molecular modeling with other residue substitutions at thenonconserved positions to probe the toxin for improvements. One canengineer changes to introduce amino acid residues with other chemicallydifferent side groups, such as opposite polarity, opposite charge, orbulky versus small.

Example 9 Focused Sequence Shuffling or Site Saturation Mutagenesis

The subject disclosure of the 3D structure of Cry 34 proteins will nowmake site- or region-directed “gene shuffling” much easier and moreefficient. U.S. Pat. No. 5,605,793, for example, describes methods forgenerating additional molecular diversity by using DNA reassembly afterrandom fragmentation. Evolutionarily conserved residues in criticalregions of the protein can now be avoided in attempting molecularevolution by shuffling or site saturation mutagenesis. This type of“shuffling” and molecular evolution can now be focused on segments, andnonconserved residues for example, in ideal regions as discussed above.

APPENDIX 1 CRYST1 100.561 100.561 56.196 90.00 90.00 90.00 I 4 2 2SCALE1 0.009944 0.000000 0.000000  0.00000 SCALE2 0.000000 0.0099440.000000  0.00000 SCALE3 0.000000 0.000000 0.017795  0.00000 ATOM 1 NALA A 3 −20.201 5.256 −9.776 1.00 30.71 N ATOM 2 CA ALA A 3 −20.3066.536 −9.021 1.00 31.43 C ATOM 3 CB ALA A 3 −21.512 6.515 −8.090 1.0030.94 C ATOM 4 C ALA A 3 −19.038 6.895 −8.247 1.00 31.22 C ATOM 5 O ALAA 3 −18.999 6.807 −7.019 1.00 31.41 O ATOM 6 N ARG A 4 −17.991 7.259−8.981 1.00 30.93 N ATOM 7 CA ARG A 4 −16.810 7.859 −8.382 1.00 30.72 CATOM 8 CB ARG A 4 −15.587 7.664 −9.303 1.00 31.38 C ATOM 9 CG ARG A 4−15.357 6.217 −9.816 1.00 32.08 C ATOM 10 CD ARG A 4 −14.418 5.337−8.967 1.00 35.83 C ATOM 11 NE ARG A 4 −14.950 3.979 −8.834 1.00 38.50 NATOM 12 CZ ARG A 4 −14.524 3.071 −7.960 1.00 41.72 C ATOM 13 NH1 ARG A 4−13.531 3.345 −7.125 1.00 41.86 N ATOM 14 NH2 ARG A 4 −15.098 1.874−7.926 1.00 41.83 N ATOM 15 C ARG A 4 −17.225 9.349 −8.279 1.00 30.38 CATOM 16 O ARG A 4 −18.053 9.783 −9.072 1.00 28.33 O ATOM 17 N GLU A 5−16.762 10.109 −7.280 1.00 30.19 N ATOM 18 CA GLU A 5 −16.964 11.581−7.297 1.00 29.80 C ATOM 19 CB GLU A 5 −18.057 12.043 −6.326 1.00 30.75C ATOM 20 CG GLU A 5 −19.037 10.967 −5.902 1.00 37.70 C ATOM 21 CD GLU A5 −19.818 11.356 −4.662 1.00 44.27 C ATOM 22 OE1 GLU A 5 −20.273 12.516−4.578 1.00 47.47 O ATOM 23 OE2 GLU A 5 −19.978 10.501 −3.770 1.00 49.26O ATOM 24 C GLU A 5 −15.634 12.299 −6.981 1.00 29.00 C ATOM 25 O GLU A 5−14.779 11.719 −6.320 1.00 29.94 O ATOM 26 N VAL A 6 −15.443 13.531−7.468 1.00 25.81 N ATOM 27 CA VAL A 6 −14.223 14.315 −7.179 1.00 25.07C ATOM 28 CB VAL A 6 −13.332 14.549 −8.419 1.00 23.55 C ATOM 29 CG1 VALA 6 −12.082 15.345 −8.037 1.00 24.08 C ATOM 30 CG2 VAL A 6 −12.94113.236 −9.051 1.00 22.59 C ATOM 31 C VAL A 6 −14.602 15.673 −6.604 1.0023.69 C ATOM 32 O VAL A 6 −15.278 16.467 −7.258 1.00 24.63 O ATOM 33 NHIS A 7 −14.185 15.927 −5.370 1.00 20.84 N ATOM 34 CA HIS A 7 −14.49417.190 −4.725 1.00 20.66 C ATOM 35 CB HIS A 7 −14.888 16.993 −3.269 1.0020.78 C ATOM 36 CG HIS A 7 −16.146 16.205 −3.080 1.00 20.91 C ATOM 37ND1 HIS A 7 −17.291 16.752 −2.542 1.00 21.69 N ATOM 38 CE1 HIS A 7−18.233 15.827 −2.483 1.00 23.78 C ATOM 39 NE2 HIS A 7 −17.738 14.699−2.959 1.00 24.50 N ATOM 40 CD2 HIS A 7 −16.434 14.908 −3.338 1.00 18.03C ATOM 41 C HIS A 7 −13.263 18.053 −4.824 1.00 20.90 C ATOM 42 O HIS A 7−12.151 17.613 −4.535 1.00 21.83 O ATOM 43 N ILE A 8 −13.469 19.295−5.226 1.00 20.16 N ATOM 44 CA ILE A 8 −12.355 20.164 −5.517 1.00 22.63C ATOM 45 CB ILE A 8 −12.343 20.450 −7.028 1.00 23.00 C ATOM 46 CG1 ILEA 8 −12.156 19.142 −7.797 1.00 23.32 C ATOM 47 CD1 ILE A 8 −12.77319.146 −9.168 1.00 23.46 C ATOM 48 CG2 ILE A 8 −11.264 21.454 −7.3841.00 23.14 C ATOM 49 C ILE A 8 −12.403 21.457 −4.741 1.00 23.55 C ATOM50 O ILE A 8 −13.375 22.206 −4.814 1.00 24.73 O ATOM 51 N ASP A 9−11.349 21.705 −3.977 1.00 21.78 N ATOM 52 CA ASP A 9 −11.206 22.976−3.300 1.00 22.57 C ATOM 53 CB ASP A 9 −10.816 22.787 −1.835 1.00 22.71C ATOM 54 CG ASP A 9 −12.010 22.500 −0.946 1.00 26.20 C ATOM 55 OD1 ASPA 9 −12.961 21.845 −1.418 1.00 27.39 O ATOM 56 OD2 ASP A 9 −12.08622.892 0.237 1.00 24.89 O ATOM 57 C ASP A 9 −10.153 23.796 −4.031 1.0023.03 C ATOM 58 O ASP A 9 −9.042 23.325 −4.279 1.00 23.74 O ATOM 59 NVAL A 10 −10.522 25.011 −4.410 1.00 22.83 N ATOM 60 CA VAL A 10 −9.58025.927 −5.025 1.00 23.02 C ATOM 61 CB VAL A 10 −10.088 26.460 −6.3781.00 24.93 C ATOM 62 CG1 VAL A 10 −9.120 27.485 −6.946 1.00 24.64 C ATOM63 CG2 VAL A 10 −10.281 25.310 −7.359 1.00 22.94 C ATOM 64 C VAL A 10−9.361 27.061 −4.032 1.00 23.75 C ATOM 65 O VAL A 10 −10.308 27.743−3.637 1.00 22.37 O ATOM 66 N ASN A 11 −8.117 27.220 −3.591 1.00 23.28 NATOM 67 CA ASN A 11 −7.761 28.273 −2.648 1.00 23.15 C ATOM 68 CB ASN A11 −7.074 27.696 −1.408 1.00 23.76 C ATOM 69 CG ASN A 11 −6.896 28.726−0.306 1.00 20.01 C ATOM 70 OD1 ASN A 11 −7.244 29.895 −0.471 1.00 21.29O ATOM 71 ND2 ASN A 11 −6.354 28.294 0.827 1.00 19.05 N ATOM 72 C ASN A11 −6.874 29.293 −3.346 1.00 23.41 C ATOM 73 O ASN A 11 −5.729 29.009−3.700 1.00 20.46 O ATOM 74 N ASN A 12 −7.429 30.482 −3.542 1.00 22.15 NATOM 75 CA ASN A 12 −6.771 31.558 −4.268 1.00 22.11 C ATOM 76 CB ASN A12 −7.845 32.400 −4.966 1.00 21.40 C ATOM 77 CG ASN A 12 −7.272 33.515−5.823 1.00 23.16 C ATOM 78 OD1 ASN A 12 −5.953 33.602 −5.892 1.00 23.90O ATOM 79 ND2 ASN A 12 −8.020 34.289 −6.422 1.00 20.48 N ATOM 80 C ASN A12 −5.926 32.445 −3.366 1.00 21.36 C ATOM 81 O ASN A 12 −6.460 33.241−2.598 1.00 20.68 O ATOM 82 N LYS A 13 −4.607 32.306 −3.453 1.00 21.06 NATOM 83 CA LYS A 13 −3.712 33.185 −2.709 1.00 21.88 C ATOM 84 CB LYS A13 −2.816 32.379 −1.759 1.00 21.33 C ATOM 85 CG LYS A 13 −3.542 31.342−0.909 1.00 25.03 C ATOM 86 CD LYS A 13 −4.573 31.977 0.009 1.00 20.87 CATOM 87 CE LYS A 13 −3.922 32.813 1.095 1.00 20.40 C ATOM 88 NZ LYS A 13−4.940 33.342 2.044 1.00 22.73 N ATOM 89 C LYS A 13 −2.840 34.024 −3.6591.00 21.42 C ATOM 90 O LYS A 13 −1.736 34.419 −3.292 1.00 21.14 O ATOM91 N THR A 14 −3.341 34.324 −4.858 1.00 22.35 N ATOM 92 CA THR A 14−2.524 34.996 −5.885 1.00 23.71 C ATOM 93 CB THR A 14 −3.071 34.723−7.305 1.00 23.77 C ATOM 94 OG1 THR A 14 −4.405 35.238 −7.417 1.00 20.92O ATOM 95 CG2 THR A 14 −3.218 33.248 −7.566 1.00 21.29 C ATOM 96 C THR A14 −2.330 36.502 −5.796 1.00 24.19 C ATOM 97 O THR A 14 −1.368 37.031−6.349 1.00 23.92 O ATOM 98 N GLY A 15 −3.250 37.204 −5.152 1.00 24.50 NATOM 99 CA GLY A 15 −3.187 38.651 −5.153 1.00 25.75 C ATOM 100 C GLY A15 −4.230 39.180 −6.119 1.00 27.59 C ATOM 101 O GLY A 15 −4.530 40.372−6.131 1.00 28.57 O ATOM 102 N HIS A 16 −4.800 38.284 −6.922 1.00 27.52N ATOM 103 CA HIS A 16 −5.810 38.679 −7.899 1.00 28.11 C ATOM 104 CB HISA 16 −5.189 38.775 −9.295 1.00 27.98 C ATOM 105 CG HIS A 16 −3.84039.423 −9.311 1.00 30.05 C ATOM 106 ND1 HIS A 16 −3.670 40.791 −9.3331.00 33.09 N ATOM 107 CE1 HIS A 16 −2.379 41.072 −9.339 1.00 34.00 CATOM 108 NE2 HIS A 16 −1.705 39.937 −9.322 1.00 32.89 N ATOM 109 CD2 HISA 16 −2.595 38.890 −9.303 1.00 28.62 C ATOM 110 C HIS A 16 −7.012 37.735−7.931 1.00 27.80 C ATOM 111 O HIS A 16 −7.064 36.747 −7.199 1.00 28.85O ATOM 112 N THR A 17 −7.979 38.060 −8.784 1.00 26.58 N ATOM 113 CA THRA 17 −9.172 37.241 −8.969 1.00 26.56 C ATOM 114 CB THR A 17 −10.32538.111 −9.507 1.00 27.04 C ATOM 115 OG1 THR A 17 −10.830 38.950 −8.4611.00 29.48 O ATOM 116 CG2 THR A 17 −11.522 37.249 −9.876 1.00 28.65 CATOM 117 C THR A 17 −8.884 36.119 −9.961 1.00 25.23 C ATOM 118 O THR A17 −8.191 36.331 −10.954 1.00 25.39 O ATOM 119 N LEU A 18 −9.390 34.927−9.678 1.00 24.46 N ATOM 120 CA LEU A 18 −9.256 33.819 −10.610 1.0024.42 C ATOM 121 CB LEU A 18 −9.209 32.475 −9.857 1.00 23.82 C ATOM 122CG LEU A 18 −7.941 32.093 −9.094 1.00 27.31 C ATOM 123 CD1 LEU A 18−8.110 30.747 −8.402 1.00 24.27 C ATOM 124 CD2 LEU A 18 −6.752 32.065−10.035 1.00 26.11 C ATOM 125 C LEU A 18 −10.419 33.778 −11.591 1.0023.64 C ATOM 126 O LEU A 18 −11.542 33.510 −11.172 1.00 22.96 O ATOM 127N GLN A 19 −10.171 34.100 −12.844 1.00 22.79 N ATOM 128 CA GLN A 19−11.257 34.088 −13.816 1.00 22.92 C ATOM 129 CB GLN A 19 −11.084 35.181−14.858 1.00 22.29 C ATOM 130 CG GLN A 19 −10.820 36.536 −14.284 1.0022.38 C ATOM 131 CD GLN A 19 −10.602 37.562 −15.364 1.00 25.21 C ATOM132 OE1 GLN A 19 −10.134 37.231 −16.453 1.00 26.18 O ATOM 133 NE2 GLN A19 −10.942 38.809 −15.076 1.00 21.40 N ATOM 134 C GLN A 19 −11.37732.758 −14.529 1.00 22.81 C ATOM 135 O GLN A 19 −10.475 32.337 −15.2511.00 24.12 O ATOM 136 N LEU A 20 −12.511 32.108 −14.327 1.00 23.53 NATOM 137 CA LEU A 20 −12.802 30.860 −14.996 1.00 24.65 C ATOM 138 CB LEUA 20 −14.196 30.404 −14.586 1.00 23.85 C ATOM 139 CG LEU A 20 −14.66729.047 −15.086 1.00 27.91 C ATOM 140 CD1 LEU A 20 −13.802 27.955 −14.4871.00 26.97 C ATOM 141 CD2 LEU A 20 −16.123 28.858 −14.712 1.00 30.71 CATOM 142 C LEU A 20 −12.759 31.069 −16.507 1.00 24.96 C ATOM 143 O LEU A20 −13.321 32.037 −17.017 1.00 25.48 O ATOM 144 N GLU A 21 −12.08430.173 −17.222 1.00 26.38 N ATOM 145 CA GLU A 21 −12.059 30.236 −18.6821.00 26.52 C ATOM 146 CB GLU A 21 −10.668 29.928 −19.217 1.00 26.85 CATOM 147 CG GLU A 21 −9.580 30.870 −18.748 1.00 34.03 C ATOM 148 CD GLUA 21 −8.240 30.479 −19.322 1.00 37.96 C ATOM 149 OE1 GLU A 21 −8.10229.305 −19.717 1.00 42.56 O ATOM 150 OE2 GLU A 21 −7.337 31.336 −19.3891.00 40.28 O ATOM 151 C GLU A 21 −13.049 29.247 −19.290 1.00 26.33 CATOM 152 O GLU A 21 −13.276 28.169 −18.742 1.00 26.34 O ATOM 153 N ASP A22 −13.616 29.611 −20.437 1.00 25.94 N ATOM 154 CA ASP A 22 −14.59428.770 −21.126 1.00 27.04 C ATOM 155 CB ASP A 22 −15.094 29.466 −22.3921.00 25.88 C ATOM 156 CG ASP A 22 −15.960 30.661 −22.091 1.00 30.27 CATOM 157 OD1 ASP A 22 −16.582 30.683 −21.009 1.00 32.86 O ATOM 158 OD2ASP A 22 −16.080 31.627 −22.873 1.00 27.23 O ATOM 159 C ASP A 22 −14.05327.401 −21.510 1.00 26.65 C ATOM 160 O ASP A 22 −14.807 26.432 −21.6021.00 26.69 O ATOM 161 N LYS A 23 −12.747 27.325 −21.741 1.00 26.01 NATOM 162 CA LYS A 23 −12.123 26.081 −22.182 1.00 26.49 C ATOM 163 CB LYSA 23 −10.722 26.345 −22.713 1.00 26.21 C ATOM 164 CG LYS A 23 −9.75326.862 −21.689 1.00 27.33 C ATOM 165 CD LYS A 23 −8.601 27.489 −22.4171.00 33.65 C ATOM 166 CE LYS A 23 −7.295 27.221 −21.733 1.00 34.50 CATOM 167 NZ LYS A 23 −6.199 27.485 −22.695 1.00 34.21 N ATOM 168 C LYS A23 −12.104 24.982 −21.122 1.00 25.01 C ATOM 169 O LYS A 23 −11.76723.836 −21.415 1.00 23.79 O ATOM 170 N THR A 24 −12.457 25.343 −19.8941.00 24.23 N ATOM 171 CA THR A 24 −12.602 24.384 −18.806 1.00 24.49 CATOM 172 CB THR A 24 −13.241 25.106 −17.608 1.00 23.78 C ATOM 173 OG1THR A 24 −12.229 25.839 −16.904 1.00 26.18 O ATOM 174 CG2 THR A 24−13.753 24.123 −16.578 1.00 24.17 C ATOM 175 C THR A 24 −13.471 23.210−19.286 1.00 25.27 C ATOM 176 O THR A 24 −14.544 23.429 −19.849 1.0024.35 O ATOM 177 N LYS A 25 −13.017 21.974 −19.074 1.00 25.29 N ATOM 178CA LYS A 25 −13.725 20.814 −19.630 1.00 27.09 C ATOM 179 CB LYS A 25−13.248 20.551 −21.075 1.00 27.03 C ATOM 180 CG LYS A 25 −13.952 19.382−21.788 1.00 30.39 C ATOM 181 CD LYS A 25 −13.731 19.362 −23.313 1.0033.98 C ATOM 182 CE LYS A 25 −12.458 18.611 −23.723 1.00 36.91 C ATOM183 NZ LYS A 25 −12.478 18.122 −25.142 1.00 33.14 N ATOM 184 C LYS A 25−13.628 19.512 −18.830 1.00 27.00 C ATOM 185 O LYS A 25 −12.599 19.208−18.225 1.00 27.27 O ATOM 186 N LEU A 26 −14.716 18.746 −18.851 1.0026.60 N ATOM 187 CA LEU A 26 −14.755 17.414 −18.258 1.00 26.61 C ATOM188 CB LEU A 26 −16.086 17.177 −17.557 1.00 27.03 C ATOM 189 CG LEU A 26−16.335 17.938 −16.264 1.00 26.27 C ATOM 190 CD1 LEU A 26 −17.671 17.522−15.693 1.00 28.25 C ATOM 191 CD2 LEU A 26 −15.219 17.659 −15.279 1.0026.42 C ATOM 192 C LEU A 26 −14.609 16.371 −19.356 1.00 27.70 C ATOM 193O LEU A 26 −15.472 16.260 −20.225 1.00 25.78 O ATOM 194 N ASP A 27−13.528 15.600 −19.320 1.00 27.94 N ATOM 195 CA ASP A 27 −13.326 14.547−20.311 1.00 29.39 C ATOM 196 CB ASP A 27 −11.876 14.078 −20.311 1.0029.51 C ATOM 197 CG ASP A 27 −10.896 15.220 −20.414 1.00 32.09 C ATOM198 OD1 ASP A 27 −10.258 15.543 −19.391 1.00 35.91 O ATOM 199 OD2 ASP A27 −10.691 15.848 −21.473 1.00 33.14 O ATOM 200 C ASP A 27 −14.23013.349 −20.047 1.00 29.05 C ATOM 201 O ASP A 27 −14.413 12.494 −20.9111.00 29.61 O ATOM 202 N GLY A 28 −14.776 13.280 −18.841 1.00 28.57 NATOM 203 CA GLY A 28 −15.651 12.190 −18.464 1.00 27.56 C ATOM 204 C GLYA 28 −16.364 12.574 −17.191 1.00 26.55 C ATOM 205 O GLY A 28 −15.75513.122 −16.275 1.00 23.93 O ATOM 206 N GLY A 29 −17.660 12.305 −17.1351.00 25.37 N ATOM 207 CA GLY A 29 −18.428 12.641 −15.957 1.00 25.02 CATOM 208 C GLY A 29 −19.185 13.938 −16.131 1.00 25.35 C ATOM 209 O GLY A29 −19.288 14.474 −17.234 1.00 25.34 O ATOM 210 N ARG A 30 −19.70114.456 −15.025 1.00 25.60 N ATOM 211 CA ARG A 30 −20.533 15.641 −15.0771.00 25.78 C ATOM 212 CB ARG A 30 −21.987 15.222 −15.230 1.00 26.40 CATOM 213 CG ARG A 30 −22.361 14.045 −14.347 1.00 27.00 C ATOM 214 CD ARGA 30 −23.710 13.439 −14.668 1.00 29.51 C ATOM 215 NE ARG A 30 −24.22612.629 −13.569 1.00 31.41 N ATOM 216 CZ ARG A 30 −25.378 11.975 −13.6121.00 35.18 C ATOM 217 NH1 ARG A 30 −26.131 12.031 −14.701 1.00 33.60 NATOM 218 NH2 ARG A 30 −25.780 11.262 −12.568 1.00 38.37 N ATOM 219 C ARGA 30 −20.373 16.501 −13.837 1.00 25.75 C ATOM 220 O ARG A 30 −19.95016.028 −12.784 1.00 26.61 O ATOM 221 N TRP A 31 −20.733 17.770 −13.9741.00 25.57 N ATOM 222 CA TRP A 31 −20.600 18.723 −12.888 1.00 25.47 CATOM 223 CB TRP A 31 −20.372 20.124 −13.457 1.00 26.48 C ATOM 224 CG TRPA 31 −19.021 20.405 −14.085 1.00 28.18 C ATOM 225 CD1 TRP A 31 −18.78320.751 −15.387 1.00 30.00 C ATOM 226 NE1 TRP A 31 −17.441 20.968 −15.5851.00 29.50 N ATOM 227 CE2 TRP A 31 −16.779 20.778 −14.400 1.00 28.44 CATOM 228 CD2 TRP A 31 −17.744 20.430 −13.430 1.00 27.86 C ATOM 229 CE3TRP A 31 −17.313 20.183 −12.122 1.00 24.54 C ATOM 230 CZ3 TRP A 31−15.962 20.288 −11.832 1.00 27.39 C ATOM 231 CH2 TRP A 31 −15.033 20.632−12.818 1.00 26.51 C ATOM 232 CZ2 TRP A 31 −15.420 20.882 −14.104 1.0027.46 C ATOM 233 C TRP A 31 −21.826 18.768 −11.977 1.00 25.20 C ATOM 234O TRP A 31 −22.926 19.107 −12.416 1.00 23.68 O ATOM 235 N ARG A 32−21.629 18.436 −10.705 1.00 24.37 N ATOM 236 CA ARG A 32 −22.691 18.535−9.715 1.00 25.26 C ATOM 237 CB ARG A 32 −22.331 17.719 −8.483 1.0024.30 C ATOM 238 CG ARG A 32 −23.541 17.197 −7.755 1.00 27.11 C ATOM 239CD ARG A 32 −23.262 16.712 −6.352 1.00 27.60 C ATOM 240 NE ARG A 32−23.046 15.271 −6.310 1.00 29.99 N ATOM 241 CZ ARG A 32 −22.676 14.606−5.228 1.00 31.20 C ATOM 242 NH1 ARG A 32 −22.472 15.249 −4.086 1.0031.03 N ATOM 243 NH2 ARG A 32 −22.507 13.293 −5.286 1.00 30.59 N ATOM244 C ARG A 32 −22.767 19.996 −9.322 1.00 25.50 C ATOM 245 O ARG A 32−23.833 20.608 −9.254 1.00 26.02 O ATOM 246 N THR A 33 −21.590 20.528−9.044 1.00 25.85 N ATOM 247 CA THR A 33 −21.410 21.909 −8.693 1.0026.91 C ATOM 248 CB THR A 33 −21.010 22.026 −7.227 1.00 27.14 C ATOM 249OG1 THR A 33 −22.098 21.590 −6.405 1.00 27.25 O ATOM 250 CG2 THR A 33−20.837 23.482 −6.839 1.00 27.63 C ATOM 251 C THR A 33 −20.280 22.280−9.609 1.00 27.22 C ATOM 252 O THR A 33 −19.229 21.639 −9.600 1.00 27.63O ATOM 253 N SER A 34 −20.500 23.284 −10.440 1.00 26.82 N ATOM 254 CASER A 34 −19.513 23.600 −11.451 1.00 26.90 C ATOM 255 CB SER A 34−20.188 23.840 −12.805 1.00 27.43 C ATOM 256 OG SER A 34 −21.241 24.778−12.697 1.00 25.73 O ATOM 257 C SER A 34 −18.634 24.763 −11.036 1.0025.93 C ATOM 258 O SER A 34 −19.037 25.610 −10.239 1.00 26.76 O ATOM 259N PRO A 35 −17.424 24.790 −11.580 1.00 25.45 N ATOM 260 CA PRO A 35−16.427 25.802 −11.224 1.00 25.90 C ATOM 261 CB PRO A 35 −15.254 25.476−12.155 1.00 26.95 C ATOM 262 CG PRO A 35 −15.448 24.054 −12.534 1.0027.75 C ATOM 263 CD PRO A 35 −16.925 23.843 −12.592 1.00 24.77 C ATOM264 C PRO A 35 −16.894 27.234 −11.467 1.00 26.85 C ATOM 265 O PRO A 35−17.629 27.492 −12.418 1.00 25.11 O ATOM 266 N THR A 36 −16.477 28.151−10.600 1.00 26.00 N ATOM 267 CA THR A 36 −16.782 29.565 −10.777 1.0026.13 C ATOM 268 CB THR A 36 −17.804 30.053 −9.739 1.00 27.21 C ATOM 269OG1 THR A 36 −17.134 30.298 −8.497 1.00 28.02 O ATOM 270 CG2 THR A 36−18.807 28.968 −9.399 1.00 30.13 C ATOM 271 C THR A 36 −15.513 30.372−10.587 1.00 25.16 C ATOM 272 O THR A 36 −14.453 29.819 −10.288 1.0025.06 O ATOM 273 N ASN A 37 −15.623 31.684 −10.763 1.00 23.04 N ATOM 274CA ASN A 37 −14.509 32.563 −10.472 1.00 23.51 C ATOM 275 CB ASN A 37−14.892 34.029 −10.673 1.00 22.69 C ATOM 276 CG ASN A 37 −15.122 34.389−12.119 1.00 23.93 C ATOM 277 OD1 ASN A 37 −14.460 33.870 −13.014 1.0021.38 O ATOM 278 ND2 ASN A 37 −16.060 35.298 −12.356 1.00 22.90 N ATOM279 C ASN A 37 −14.241 32.381 −9.001 1.00 22.99 C ATOM 280 O ASN A 37−15.136 31.998 −8.250 1.00 21.76 O ATOM 281 N VAL A 38 −13.019 32.632−8.566 1.00 21.81 N ATOM 282 CA VAL A 38 −12.807 32.648 −7.132 1.0023.03 C ATOM 283 CB VAL A 38 −12.122 31.358 −6.649 1.00 23.82 C ATOM 284CG1 VAL A 38 −11.799 31.449 −5.165 1.00 26.55 C ATOM 285 CG2 VAL A 38−12.998 30.149 −6.937 1.00 22.08 C ATOM 286 C VAL A 38 −11.971 33.841−6.731 1.00 22.62 C ATOM 287 O VAL A 38 −10.937 34.150 −7.330 1.00 22.28O ATOM 288 N ALA A 39 −12.481 34.529 −5.721 1.00 22.00 N ATOM 289 CA ALAA 39 −11.898 35.752 −5.210 1.00 24.24 C ATOM 290 CB ALA A 39 −12.88336.412 −4.259 1.00 23.20 C ATOM 291 C ALA A 39 −10.584 35.515 −4.4941.00 25.00 C ATOM 292 O ALA A 39 −10.342 34.433 −3.958 1.00 24.33 O ATOM293 N ASN A 40 −9.735 36.536 −4.484 1.00 25.96 N ATOM 294 CA ASN A 40−8.518 36.461 −3.698 1.00 26.45 C ATOM 295 CB ASN A 40 −7.735 37.771−3.746 1.00 25.97 C ATOM 296 CG ASN A 40 −6.373 37.650 −3.090 1.00 28.17C ATOM 297 OD1 ASN A 40 −5.633 36.612 −3.461 1.00 25.91 O ATOM 298 ND2ASN A 40 −5.995 38.469 −2.252 1.00 26.68 N ATOM 299 C ASN A 40 −8.88536.128 −2.256 1.00 26.05 C ATOM 300 O ASN A 40 −9.948 36.524 −1.767 1.0023.60 O ATOM 301 N ASP A 41 −7.998 35.392 −1.594 1.00 25.12 N ATOM 302CA ASP A 41 −8.187 34.949 −0.212 1.00 23.72 C ATOM 303 CB ASP A 41−8.005 36.100 0.774 1.00 23.36 C ATOM 304 CG ASP A 41 −6.611 36.6720.740 1.00 28.68 C ATOM 305 OD1 ASP A 41 −5.650 35.892 0.570 1.00 24.35O ATOM 306 OD2 ASP A 41 −6.381 37.891 0.867 1.00 26.98 O ATOM 307 C ASPA 41 −9.530 34.280 0.013 1.00 22.57 C ATOM 308 O ASP A 41 −10.162 34.4621.052 1.00 22.71 O ATOM 309 N GLN A 42 −9.955 33.496 −0.966 1.00 22.25 NATOM 310 CA GLN A 42 −11.215 32.786 −0.870 1.00 22.19 C ATOM 311 CB GLNA 42 −12.280 33.467 −1.747 1.00 21.73 C ATOM 312 CG GLN A 42 −13.52032.623 −2.055 1.00 21.24 C ATOM 313 CD GLN A 42 −14.591 33.384 −2.8331.00 23.32 C ATOM 314 OE1 GLN A 42 −14.665 33.287 −4.060 1.00 23.60 OATOM 315 NE2 GLN A 42 −15.429 34.128 −2.118 1.00 17.55 N ATOM 316 C GLNA 42 −11.021 31.332 −1.274 1.00 21.63 C ATOM 317 O GLN A 42 −10.13431.006 −2.067 1.00 19.50 O ATOM 318 N ILE A 43 −11.834 30.462 −0.6851.00 22.96 N ATOM 319 CA ILE A 43 −11.865 29.054 −1.045 1.00 24.37 CATOM 320 CB ILE A 43 −11.506 28.158 0.154 1.00 26.25 C ATOM 321 CG1 ILEA 43 −10.135 28.514 0.721 1.00 28.14 C ATOM 322 CD1 ILE A 43 −9.86327.871 2.063 1.00 31.84 C ATOM 323 CG2 ILE A 43 −11.549 26.688 −0.2491.00 25.52 C ATOM 324 C ILE A 43 −13.275 28.704 −1.497 1.00 24.15 C ATOM325 O ILE A 43 −14.253 29.072 −0.845 1.00 22.87 O ATOM 326 N LYS A 44−13.374 28.010 −2.623 1.00 22.89 N ATOM 327 CA LYS A 44 −14.653 27.515−3.103 1.00 24.30 C ATOM 328 CB LYS A 44 −15.131 28.248 −4.360 1.0024.88 C ATOM 329 CG LYS A 44 −15.702 29.623 −4.069 1.00 25.52 C ATOM 330CD LYS A 44 −17.043 29.857 −4.754 1.00 29.82 C ATOM 331 CE LYS A 44−16.905 30.758 −5.962 1.00 34.24 C ATOM 332 NZ LYS A 44 −18.014 31.749−6.025 1.00 41.35 N ATOM 333 C LYS A 44 −14.508 26.035 −3.375 1.00 24.64C ATOM 334 O LYS A 44 −13.402 25.530 −3.574 1.00 24.26 O ATOM 335 N THRA 45 −15.634 25.343 −3.378 1.00 23.64 N ATOM 336 CA THR A 45 −15.63823.912 −3.571 1.00 23.75 C ATOM 337 CB THR A 45 −16.227 23.227 −2.3291.00 24.63 C ATOM 338 OG1 THR A 45 −15.438 23.564 −1.180 1.00 25.23 OATOM 339 CG2 THR A 45 −16.088 21.716 −2.425 1.00 22.69 C ATOM 340 C THRA 45 −16.479 23.607 −4.788 1.00 24.16 C ATOM 341 O THR A 45 −17.59524.101 −4.921 1.00 23.69 O ATOM 342 N PHE A 46 −15.929 22.814 −5.6931.00 24.62 N ATOM 343 CA PHE A 46 −16.668 22.410 −6.872 1.00 24.00 CATOM 344 CB PHE A 46 −15.966 22.917 −8.126 1.00 24.93 C ATOM 345 CG PHEA 46 −15.660 24.389 −8.092 1.00 26.04 C ATOM 346 CD1 PHE A 46 −14.38024.850 −8.350 1.00 29.54 C ATOM 347 CE1 PHE A 46 −14.095 26.201 −8.3201.00 26.62 C ATOM 348 CZ PHE A 46 −15.092 27.110 −8.028 1.00 26.20 CATOM 349 CE2 PHE A 46 −16.373 26.666 −7.766 1.00 26.71 C ATOM 350 CD2PHE A 46 −16.652 25.312 −7.798 1.00 24.99 C ATOM 351 C PHE A 46 −16.73320.896 −6.842 1.00 23.71 C ATOM 352 O PHE A 46 −15.910 20.261 −6.1841.00 22.84 O ATOM 353 N VAL A 47 −17.712 20.306 −7.519 1.00 23.04 N ATOM354 CA VAL A 47 −17.822 18.848 −7.505 1.00 24.28 C ATOM 355 CB VAL A 47−18.847 18.349 −6.466 1.00 25.54 C ATOM 356 CG1 VAL A 47 −18.621 16.869−6.193 1.00 24.52 C ATOM 357 CG2 VAL A 47 −18.770 19.151 −5.179 1.0022.94 C ATOM 358 C VAL A 47 −18.209 18.190 −8.825 1.00 25.74 C ATOM 359O VAL A 47 −19.154 18.610 −9.492 1.00 24.05 O ATOM 360 N ALA A 48−17.502 17.113 −9.157 1.00 26.98 N ATOM 361 CA ALA A 48 −17.764 16.357−10.377 1.00 29.96 C ATOM 362 CB ALA A 48 −16.533 16.368 −11.267 1.0029.88 C ATOM 363 C ALA A 48 −18.223 14.916 −10.106 1.00 30.80 C ATOM 364O ALA A 48 −17.763 14.275 −9.159 1.00 32.03 O ATOM 365 N GLU A 49−19.118 14.415 −10.958 1.00 30.76 N ATOM 366 CA GLU A 49 −19.688 13.070−10.827 1.00 32.41 C ATOM 367 CB GLU A 49 −21.208 13.165 −10.674 1.0032.52 C ATOM 368 CG GLU A 49 −21.685 13.750 −9.361 1.00 33.36 C ATOM 369CD GLU A 49 −23.135 14.190 −9.422 1.00 35.31 C ATOM 370 OE1 GLU A 49−23.631 14.468 −10.535 1.00 39.64 O ATOM 371 OE2 GLU A 49 −23.778 14.262−8.356 1.00 30.65 O ATOM 372 C GLU A 49 −19.405 12.167 −12.028 1.0032.60 C ATOM 373 O GLU A 49 −19.627 12.566 −13.169 1.00 31.95 O ATOM 374N SER A 50 −18.921 10.953 −11.775 1.00 32.73 N ATOM 375 CA SER A 50−18.743 9.971 −12.843 1.00 32.17 C ATOM 376 CB SER A 50 −17.820 8.837−12.391 1.00 32.15 C ATOM 377 OG SER A 50 −18.278 7.582 −12.862 1.0028.65 O ATOM 378 C SER A 50 −20.151 9.457 −13.137 1.00 33.46 C ATOM 379O SER A 50 −20.925 9.249 −12.207 1.00 32.67 O ATOM 380 N ASN A 51−20.500 9.270 −14.409 1.00 34.65 N ATOM 381 CA ASN A 51 −21.875 8.906−14.756 1.00 36.67 C ATOM 382 CB ASN A 51 −22.574 10.121 −15.376 1.0038.31 C ATOM 383 CG ASN A 51 −22.016 10.479 −16.745 1.00 43.96 C ATOM384 OD1 ASN A 51 −21.146 9.788 −17.271 1.00 48.46 O ATOM 385 ND2 ASN A51 −22.519 11.562 −17.327 1.00 48.63 N ATOM 386 C ASN A 51 −22.091 7.690−15.672 1.00 35.67 C ATOM 387 O ASN A 51 −23.168 7.545 −16.257 1.0036.15 O ATOM 388 N GLY A 52 −21.094 6.821 −15.801 1.00 33.90 N ATOM 389CA GLY A 52 −21.223 5.665 −16.680 1.00 32.29 C ATOM 390 C GLY A 52−20.558 4.367 −16.241 1.00 30.58 C ATOM 391 O GLY A 52 −19.988 4.284−15.154 1.00 30.65 O ATOM 392 N PHE A 53 −20.626 3.355 −17.108 1.0028.54 N ATOM 393 CA PHE A 53 −20.078 2.019 −16.836 1.00 26.49 C ATOM 394CB PHE A 53 −20.776 0.981 −17.717 1.00 26.00 C ATOM 395 CG PHE A 53−20.607 −0.444 −17.249 1.00 24.59 C ATOM 396 CD1 PHE A 53 −20.689 −0.766−15.904 1.00 23.57 C ATOM 397 CE1 PHE A 53 −20.554 −2.077 −15.478 1.0021.50 C ATOM 398 CZ PHE A 53 −20.337 −3.083 −16.399 1.00 21.31 C ATOM399 CE2 PHE A 53 −20.257 −2.778 −17.742 1.00 22.36 C ATOM 400 CD2 PHE A53 −20.394 −1.466 −18.162 1.00 23.09 C ATOM 401 C PHE A 53 −18.585 1.927−17.104 1.00 25.97 C ATOM 402 O PHE A 53 −18.130 2.227 −18.206 1.0025.48 O ATOM 403 N MET A 54 −17.831 1.477 −16.107 1.00 25.47 N ATOM 404CA MET A 54 −16.384 1.339 −16.242 1.00 25.06 C ATOM 405 CB MET A 54−16.026 0.160 −17.148 1.00 26.08 C ATOM 406 CG MET A 54 −16.471 −1.196−16.649 1.00 27.87 C ATOM 407 SD MET A 54 −15.754 −2.520 −17.642 1.0035.01 S ATOM 408 CE MET A 54 −15.698 −1.744 −19.257 1.00 35.63 C ATOM409 C MET A 54 −15.714 2.590 −16.797 1.00 24.23 C ATOM 410 O MET A 54−14.777 2.493 −17.585 1.00 24.08 O ATOM 411 N THR A 55 −16.223 3.759−16.435 1.00 23.87 N ATOM 412 CA THR A 55 −15.563 5.004 −16.793 1.0023.80 C ATOM 413 CB THR A 55 −16.381 5.876 −17.761 1.00 24.73 C ATOM 414OG1 THR A 55 −17.738 5.961 −17.307 1.00 26.42 O ATOM 415 CG2 THR A 55−16.481 5.246 −19.138 1.00 25.56 C ATOM 416 C THR A 55 −15.439 5.751−15.501 1.00 23.22 C ATOM 417 O THR A 55 −15.879 5.280 −14.454 1.0022.19 O ATOM 418 N GLY A 56 −14.875 6.942 −15.575 1.00 25.23 N ATOM 419CA GLY A 56 −14.687 7.712 −14.374 1.00 25.93 C ATOM 420 C GLY A 56−14.827 9.188 −14.613 1.00 27.72 C ATOM 421 O GLY A 56 −15.301 9.638−15.658 1.00 28.31 O ATOM 422 N THR A 57 −14.409 9.944 −13.613 1.0027.41 N ATOM 423 CA THR A 57 −14.454 11.382 −13.687 1.00 28.12 C ATOM424 CB THR A 57 −14.827 11.963 −12.328 1.00 28.49 C ATOM 425 OG1 THR A57 −16.192 11.658 −12.027 1.00 28.54 O ATOM 426 CG2 THR A 57 −14.80713.475 −12.398 1.00 29.80 C ATOM 427 C THR A 57 −13.088 11.906 −14.0481.00 28.27 C ATOM 428 O THR A 57 −12.095 11.553 −13.414 1.00 27.85 OATOM 429 N GLU A 58 −13.028 12.776 −15.043 1.00 26.04 N ATOM 430 CA GLUA 58 −11.747 13.345 −15.402 1.00 25.04 C ATOM 431 CB GLU A 58 −10.98612.369 −16.287 1.00 24.49 C ATOM 432 CG GLU A 58 −9.744 12.957 −16.9191.00 26.73 C ATOM 433 CD GLU A 58 −9.131 12.007 −17.917 1.00 32.10 CATOM 434 OE1 GLU A 58 −9.867 11.134 −18.423 1.00 31.45 O ATOM 435 OE2GLU A 58 −7.920 12.129 −18.189 1.00 34.10 O ATOM 436 C GLU A 58 −11.86514.688 −16.097 1.00 25.39 C ATOM 437 O GLU A 58 −12.619 14.836 −17.0561.00 23.42 O ATOM 438 N GLY A 59 −11.106 15.667 −15.620 1.00 24.93 NATOM 439 CA GLY A 59 −11.136 16.968 −16.251 1.00 24.69 C ATOM 440 C GLYA 59 −10.050 17.954 −15.882 1.00 24.92 C ATOM 441 O GLY A 59 −9.12617.660 −15.118 1.00 23.09 O ATOM 442 N THR A 60 −10.190 19.147 −16.4491.00 23.57 N ATOM 443 CA THR A 60 −9.256 20.236 −16.240 1.00 24.21 CATOM 444 CB THR A 60 −8.290 20.344 −17.429 1.00 25.09 C ATOM 445 OG1 THRA 60 −7.624 19.092 −17.630 1.00 24.87 O ATOM 446 CG2 THR A 60 −7.15921.308 −17.106 1.00 24.43 C ATOM 447 C THR A 60 −9.996 21.552 −16.1161.00 24.54 C ATOM 448 O THR A 60 −10.766 21.928 −17.000 1.00 25.46 OATOM 449 N ILE A 61 −9.762 22.251 −15.014 1.00 23.54 N ATOM 450 CA ILE A61 −10.325 23.577 −14.837 1.00 23.91 C ATOM 451 CB ILE A 61 −10.72623.817 −13.383 1.00 24.15 C ATOM 452 CG1 ILE A 61 −11.749 22.785 −12.9171.00 23.08 C ATOM 453 CD1 ILE A 61 −12.049 22.881 −11.439 1.00 18.53 CATOM 454 CG2 ILE A 61 −11.274 25.226 −13.222 1.00 23.66 C ATOM 455 C ILEA 61 −9.256 24.586 −15.204 1.00 24.81 C ATOM 456 O ILE A 61 −8.12924.506 −14.715 1.00 25.35 O ATOM 457 N TYR A 62 −9.608 25.542 −16.0541.00 24.24 N ATOM 458 CA TYR A 62 −8.668 26.580 −16.445 1.00 24.50 CATOM 459 CB TYR A 62 −8.594 26.693 −17.960 1.00 24.65 C ATOM 460 CG TYRA 62 −8.051 25.487 −18.680 1.00 26.90 C ATOM 461 CD1 TYR A 62 −8.90724.546 −19.229 1.00 25.16 C ATOM 462 CE1 TYR A 62 −8.420 23.451 −19.9091.00 27.18 C ATOM 463 CZ TYR A 62 −7.060 23.290 −20.054 1.00 26.12 CATOM 464 OH TYR A 62 −6.573 22.197 −20.731 1.00 26.72 O ATOM 465 CE2 TYRA 62 −6.186 24.216 −19.523 1.00 24.12 C ATOM 466 CD2 TYR A 62 −6.68425.309 −18.844 1.00 25.80 C ATOM 467 C TYR A 62 −9.046 27.948 −15.8991.00 24.69 C ATOM 468 O TYR A 62 −10.178 28.406 −16.068 1.00 25.15 OATOM 469 N TYR A 63 −8.082 28.609 −15.269 1.00 23.33 N ATOM 470 CA TYR A63 −8.287 29.955 −14.760 1.00 24.10 C ATOM 471 CB TYR A 63 −8.147 30.004−13.241 1.00 24.17 C ATOM 472 CG TYR A 63 −9.302 29.446 −12.449 1.0026.52 C ATOM 473 CD1 TYR A 63 −9.170 28.252 −11.759 1.00 27.18 C ATOM474 CE1 TYR A 63 −10.208 27.737 −11.016 1.00 28.68 C ATOM 475 CZ TYR A63 −11.400 28.422 −10.947 1.00 24.41 C ATOM 476 OH TYR A 63 −12.43627.906 −10.205 1.00 18.74 O ATOM 477 CE2 TYR A 63 −11.558 29.619 −11.6141.00 25.27 C ATOM 478 CD2 TYR A 63 −10.509 30.128 −12.356 1.00 27.38 CATOM 479 C TYR A 63 −7.269 30.921 −15.338 1.00 24.54 C ATOM 480 O TYR A63 −6.135 30.551 −15.649 1.00 23.25 O ATOM 481 N SER A 64 −7.690 32.174−15.434 1.00 25.09 N ATOM 482 CA SER A 64 −6.856 33.260 −15.905 1.0025.59 C ATOM 483 CB SER A 64 −7.612 34.025 −16.994 1.00 24.67 C ATOM 484OG SER A 64 −6.897 35.162 −17.439 1.00 24.48 O ATOM 485 C SER A 64−6.554 34.198 −14.736 1.00 27.22 C ATOM 486 O SER A 64 −7.433 34.480−13.923 1.00 27.09 O ATOM 487 N ILE A 65 −5.309 34.651 −14.627 1.0029.47 N ATOM 488 CA ILE A 65 −4.967 35.674 −13.646 1.00 33.07 C ATOM 489CB ILE A 65 −3.669 35.308 −12.873 1.00 32.14 C ATOM 490 CG1 ILE A 65−3.859 33.978 −12.131 1.00 32.81 C ATOM 491 CD1 ILE A 65 −2.592 33.411−11.519 1.00 32.19 C ATOM 492 CG2 ILE A 65 −3.294 36.399 −11.879 1.0035.96 C ATOM 493 C ILE A 65 −4.878 36.998 −14.424 1.00 34.72 C ATOM 494O ILE A 65 −3.849 37.310 −15.024 1.00 34.58 O ATOM 495 N ASN A 66 −6.02537.683 −14.503 1.00 38.05 N ATOM 496 CA ASN A 66 −6.179 39.056 −15.0181.00 39.99 C ATOM 497 CB ASN A 66 −5.946 40.075 −13.903 1.00 42.08 CATOM 498 CG ASN A 66 −6.920 39.913 −12.752 1.00 47.88 C ATOM 499 OD1 ASNA 66 −8.104 39.654 −12.961 1.00 54.24 O ATOM 500 ND2 ASN A 66 −6.41840.044 −11.531 1.00 52.65 N ATOM 501 C ASN A 66 −5.246 39.498 −16.1471.00 39.26 C ATOM 502 O ASN A 66 −4.584 40.537 −16.052 1.00 39.80 O ATOM503 N GLY A 67 −5.190 38.689 −17.201 1.00 38.73 N ATOM 504 CA GLY A 67−4.446 39.023 −18.405 1.00 37.19 C ATOM 505 C GLY A 67 −2.989 38.650−18.300 1.00 36.09 C ATOM 506 O GLY A 67 −2.225 38.720 −19.265 1.0037.58 O ATOM 507 N GLU A 68 −2.629 38.204 −17.108 1.00 33.60 N ATOM 508CA GLU A 68 −1.256 37.894 −16.772 1.00 32.14 C ATOM 509 CB GLU A 68−0.993 38.393 −15.349 1.00 33.24 C ATOM 510 CG GLU A 68 0.461 38.592−14.986 1.00 35.30 C ATOM 511 CD GLU A 68 0.676 38.588 −13.484 1.0038.70 C ATOM 512 OE1 GLU A 68 −0.277 38.717 −12.714 1.00 38.53 O ATOM513 OE2 GLU A 68 1.925 38.431 −13.064 1.00 37.16 N ATOM 514 C GLU A 68−0.821 36.424 −16.892 1.00 29.98 C ATOM 515 O GLU A 68 0.318 36.161−17.268 1.00 28.96 O ATOM 516 N ALA A 69 −1.690 35.467 −16.570 1.0026.86 N ATOM 517 CA ALA A 69 −1.254 34.066 −16.583 1.00 25.20 C ATOM 518CB ALA A 69 −0.341 33.795 −15.389 1.00 24.76 C ATOM 519 C ALA A 69−2.375 33.037 −16.611 1.00 24.66 C ATOM 520 O ALA A 69 −3.491 33.302−16.170 1.00 23.50 O ATOM 521 N GLU A 70 −2.058 31.848 −17.113 1.0023.75 N ATOM 522 CA GLU A 70 −3.035 30.770 −17.158 1.00 24.20 C ATOM 523CB GLU A 70 −3.127 30.166 −18.560 1.00 23.76 C ATOM 524 CG GLU A 70−4.470 29.502 −18.803 1.00 26.21 C ATOM 525 CD GLU A 70 −4.569 28.786−20.133 1.00 30.59 C ATOM 526 OE1 GLU A 70 −3.638 28.036 −20.493 1.0035.67 O ATOM 527 OE2 GLU A 70 −5.600 28.964 −20.810 1.00 28.27 O ATOM528 C GLU A 70 −2.739 29.673 −16.135 1.00 24.75 C ATOM 529 O GLU A 70−1.606 29.202 −16.022 1.00 23.63 O ATOM 530 N ILE A 71 −3.765 29.281−15.385 1.00 23.84 N ATOM 531 CA ILE A 71 −3.649 28.179 −14.436 1.0025.53 C ATOM 532 CB ILE A 71 −4.204 28.578 −13.054 1.00 24.67 C ATOM 533CG1 ILE A 71 −3.591 29.893 −12.571 1.00 28.07 C ATOM 534 CD1 ILE A 71−2.113 29.808 −12.278 1.00 29.93 C ATOM 535 CG2 ILE A 71 −3.965 27.460−12.047 1.00 25.65 C ATOM 536 C ILE A 71 −4.428 26.968 −14.943 1.0025.20 C ATOM 537 O ILE A 71 −5.555 27.108 −15.418 1.00 24.13 O ATOM 538N SER A 72 −3.831 25.783 −14.842 1.00 23.65 N ATOM 539 CA SER A 72−4.509 24.548 −15.227 1.00 22.64 C ATOM 540 CB SER A 72 −3.801 23.869−16.400 1.00 22.07 C ATOM 541 OG SER A 72 −3.530 24.784 −17.443 1.0027.15 O ATOM 542 C SER A 72 −4.564 23.580 −14.051 1.00 21.99 C ATOM 543O SER A 72 −3.536 23.249 −13.461 1.00 21.52 O ATOM 544 N LEU A 73 −5.76823.128 −13.717 1.00 22.03 N ATOM 545 CA LEU A 73 −5.956 22.188 −12.6201.00 21.77 C ATOM 546 CB LEU A 73 −6.813 22.808 −11.513 1.00 21.46 CATOM 547 CG LEU A 73 −6.474 24.245 −11.090 1.00 22.77 C ATOM 548 CD1 LEUA 73 −7.560 24.812 −10.183 1.00 23.15 C ATOM 549 CD2 LEU A 73 −5.10224.346 −10.424 1.00 16.28 C ATOM 550 C LEU A 73 −6.594 20.921 −13.1731.00 23.85 C ATOM 551 O LEU A 73 −7.752 20.921 −13.593 1.00 22.12 O ATOM552 N TYR A 74 −5.813 19.848 −13.183 1.00 22.69 N ATOM 553 CA TYR A 74−6.239 18.567 −13.728 1.00 22.49 C ATOM 554 CB TYR A 74 −5.120 17.995−14.612 1.00 21.92 C ATOM 555 CG TYR A 74 −5.304 16.565 −15.085 1.0022.39 C ATOM 556 CD1 TYR A 74 −6.152 16.263 −16.143 1.00 25.97 C ATOM557 CE1 TYR A 74 −6.313 14.959 −16.584 1.00 27.83 C ATOM 558 CZ TYR A 74−5.614 13.940 −15.971 1.00 29.25 C ATOM 559 OH TYR A 74 −5.773 12.644−16.410 1.00 25.74 O ATOM 560 CE2 TYR A 74 −4.759 14.214 −14.924 1.0025.79 C ATOM 561 CD2 TYR A 74 −4.603 15.520 −14.491 1.00 23.40 C ATOM562 C TYR A 74 −6.579 17.597 −12.608 1.00 22.85 C ATOM 563 O TYR A 74−5.907 17.562 −11.578 1.00 21.60 O ATOM 564 N PHE A 75 −7.644 16.830−12.803 1.00 21.68 N ATOM 565 CA PHE A 75 −8.019 15.793 −11.854 1.0022.34 C ATOM 566 CB PHE A 75 −9.126 16.269 −10.912 1.00 22.81 C ATOM 567CG PHE A 75 −10.344 16.781 −11.620 1.00 23.45 C ATOM 568 CD1 PHE A 75−11.353 15.917 −12.017 1.00 23.17 C ATOM 569 CE1 PHE A 75 −12.471 16.393−12.673 1.00 27.45 C ATOM 570 CZ PHE A 75 −12.590 17.743 −12.936 1.0022.60 C ATOM 571 CE2 PHE A 75 −11.592 18.611 −12.547 1.00 23.01 C ATOM572 CD2 PHE A 75 −10.479 18.130 −11.893 1.00 23.73 C ATOM 573 C PHE A 75−8.474 14.552 −12.605 1.00 21.45 C ATOM 574 O PHE A 75 −9.048 14.644−13.691 1.00 20.44 O ATOM 575 N ASP A 76 −8.199 13.392 −12.021 1.0020.36 N ATOM 576 CA ASP A 76 −8.636 12.123 −12.584 1.00 19.20 C ATOM 577CB ASP A 76 −7.596 11.561 −13.556 1.00 19.48 C ATOM 578 CG ASP A 76−8.096 10.335 −14.302 1.00 22.24 C ATOM 579 OD1 ASP A 76 −9.329 10.155−14.402 1.00 21.23 O ATOM 580 OD2 ASP A 76 −7.330 9.497 −14.822 1.0028.22 O ATOM 581 C ASP A 76 −8.921 11.125 −11.467 1.00 18.15 C ATOM 582O ASP A 76 −8.063 10.842 −10.633 1.00 15.04 O ATOM 583 N ASN A 77−10.149 10.622 −11.450 1.00 16.70 N ATOM 584 CA ASN A 77 −10.593 9.616−10.501 1.00 17.65 C ATOM 585 CB ASN A 77 −11.787 10.155 −9.703 1.0016.46 C ATOM 586 CG ASN A 77 −12.111 9.327 −8.463 1.00 20.79 C ATOM 587OD1 ASN A 77 −11.996 8.102 −8.463 1.00 15.07 O ATOM 588 ND2 ASN A 77−12.539 10.005 −7.401 1.00 18.53 N ATOM 589 C ASN A 77 −11.029 8.485−11.412 1.00 20.12 C ATOM 590 O ASN A 77 −12.203 8.401 −11.771 1.0020.56 O ATOM 591 N PRO A 78 −10.091 7.647 −11.846 1.00 20.61 N ATOM 592CA PRO A 78 −10.442 6.618 −12.821 1.00 20.41 C ATOM 593 CB PRO A 78−9.094 6.059 −13.296 1.00 21.13 C ATOM 594 CG PRO A 78 −8.047 6.711−12.501 1.00 19.86 C ATOM 595 CD PRO A 78 −8.670 7.594 −11.464 1.0019.47 C ATOM 596 C PRO A 78 −11.260 5.500 −12.252 1.00 22.03 C ATOM 597O PRO A 78 −11.557 5.402 −11.058 1.00 20.62 O ATOM 598 N PHE A 79−11.601 4.623 −13.174 1.00 22.52 N ATOM 599 CA PHE A 79 −12.444 3.504−12.890 1.00 23.95 C ATOM 600 CB PHE A 79 −12.727 2.777 −14.177 1.0023.84 C ATOM 601 CG PHE A 79 −13.351 1.491 −13.946 1.00 22.20 C ATOM 602CD1 PHE A 79 −14.346 1.399 −13.001 1.00 21.49 C ATOM 603 CE1 PHE A 79−14.930 0.216 −12.725 1.00 21.53 C ATOM 604 CZ PHE A 79 −14.516 −0.914−13.386 1.00 22.05 C ATOM 605 CE2 PHE A 79 −13.503 −0.840 −14.322 1.0020.33 C ATOM 606 CD2 PHE A 79 −12.917 0.360 −14.591 1.00 20.10 C ATOM607 C PHE A 79 −11.774 2.564 −11.900 1.00 25.71 C ATOM 608 O PHE A 79−12.321 2.243 −10.846 1.00 25.70 O ATOM 609 N ALA A 80 −10.602 2.084−12.275 1.00 26.68 N ATOM 610 CA ALA A 80 −9.767 1.353 −11.353 1.0028.43 C ATOM 611 CB ALA A 80 −9.698 −0.099 −11.710 1.00 28.35 C ATOM 612C ALA A 80 −8.435 2.024 −11.559 1.00 29.41 C ATOM 613 O ALA A 80 −8.0742.364 −12.685 1.00 30.73 O ATOM 614 N GLY A 81 −7.714 2.261 −10.480 1.0029.53 N ATOM 615 CA GLY A 81 −6.415 2.867 −10.619 1.00 29.20 C ATOM 616C GLY A 81 −6.176 3.856 −9.520 1.00 29.47 C ATOM 617 O GLY A 81 −6.9233.929 −8.545 1.00 30.93 O ATOM 618 N SER A 82 −5.102 4.607 −9.659 1.0027.97 N ATOM 619 CA SER A 82 −4.851 5.640 −8.700 1.00 26.30 C ATOM 620CB SER A 82 −3.355 5.795 −8.445 1.00 26.99 C ATOM 621 OG SER A 82 −2.7844.554 −8.071 1.00 30.52 O ATOM 622 C SER A 82 −5.423 6.903 −9.293 1.0024.01 C ATOM 623 O SER A 82 −5.406 7.102 −10.511 1.00 21.96 O ATOM 624 NASN A 83 −5.986 7.725 −8.424 1.00 22.72 N ATOM 625 CA ASN A 83 −6.4099.051 −8.802 1.00 23.89 C ATOM 626 CB ASN A 83 −7.070 9.724 −7.601 1.0023.53 C ATOM 627 CG ASN A 83 −8.403 9.093 −7.235 1.00 28.45 C ATOM 628OD1 ASN A 83 −9.183 8.720 −8.107 1.00 29.35 O ATOM 629 ND2 ASN A 83−8.674 8.982 −5.939 1.00 31.22 N ATOM 630 C ASN A 83 −5.162 9.830 −9.2321.00 23.74 C ATOM 631 O ASN A 83 −4.062 9.564 −8.744 1.00 23.77 O ATOM632 N LYS A 84 −5.323 10.767 −10.161 1.00 23.79 N ATOM 633 CA LYS A 84−4.209 11.591 −10.626 1.00 24.20 C ATOM 634 CB LYS A 84 −3.842 11.264−12.079 1.00 25.35 C ATOM 635 CG LYS A 84 −3.097 9.958 −12.270 1.0028.77 C ATOM 636 CD LYS A 84 −2.470 9.851 −13.661 1.00 36.11 C ATOM 637CE LYS A 84 −3.485 9.429 −14.707 1.00 40.41 C ATOM 638 NZ LYS A 84−2.918 8.424 −15.653 1.00 44.23 N ATOM 639 C LYS A 84 −4.603 13.051−10.537 1.00 23.62 C ATOM 640 O LYS A 84 −5.752 13.403 −10.793 1.0021.79 O ATOM 641 N TYR A 85 −3.657 13.905 −10.168 1.00 22.99 N ATOM 642CA TYR A 85 −3.936 15.331 −10.108 1.00 23.66 C ATOM 643 CB TYR A 85−4.434 15.748 −8.718 1.00 22.81 C ATOM 644 CG TYR A 85 −5.471 14.872−8.029 1.00 25.71 C ATOM 645 CD1 TYR A 85 −5.091 13.773 −7.266 1.0028.50 C ATOM 646 CE1 TYR A 85 −6.032 12.994 −6.607 1.00 28.72 C ATOM 647CZ TYR A 85 −7.370 13.323 −6.691 1.00 27.90 C ATOM 648 OH TYR A 85−8.314 12.558 −6.041 1.00 26.65 O ATOM 649 CE2 TYR A 85 −7.770 14.417−7.426 1.00 26.22 C ATOM 650 CD2 TYR A 85 −6.824 15.190 −8.082 1.0024.50 C ATOM 651 C TYR A 85 −2.690 16.155 −10.433 1.00 24.09 C ATOM 652O TYR A 85 −1.567 15.734 −10.151 1.00 21.65 O ATOM 653 N ASP A 86 −2.89717.324 −11.034 1.00 23.43 N ATOM 654 CA ASP A 86 −1.814 18.283 −11.2541.00 24.93 C ATOM 655 CB ASP A 86 −0.979 17.961 −12.495 1.00 24.97 CATOM 656 CG ASP A 86 0.474 18.390 −12.341 1.00 27.97 C ATOM 657 OD1 ASPA 86 0.861 18.775 −11.216 1.00 26.47 O ATOM 658 OD2 ASP A 86 1.30318.367 −13.275 1.00 30.47 O ATOM 659 C ASP A 86 −2.340 19.708 −11.3501.00 24.39 C ATOM 660 O ASP A 86 −3.472 19.944 −11.773 1.00 25.08 O ATOM661 N GLY A 87 −1.503 20.650 −10.935 1.00 25.42 N ATOM 662 CA GLY A 87−1.804 22.061 −11.036 1.00 26.07 C ATOM 663 C GLY A 87 −0.607 22.726−11.686 1.00 27.17 C ATOM 664 O GLY A 87 0.523 22.534 −11.239 1.00 25.48O ATOM 665 N HIS A 88 −0.837 23.498 −12.742 1.00 26.94 N ATOM 666 CA HISA 88 0.274 24.131 −13.447 1.00 27.76 C ATOM 667 CB HIS A 88 0.659 23.299−14.674 1.00 27.46 C ATOM 668 CG HIS A 88 1.617 23.981 −15.603 1.0030.53 C ATOM 669 ND1 HIS A 88 2.974 23.739 −15.583 1.00 30.43 N ATOM 670CE1 HIS A 88 3.563 24.465 −16.517 1.00 29.34 C ATOM 671 NE2 HIS A 882.636 25.166 −17.145 1.00 30.58 N ATOM 672 CD2 HIS A 88 1.410 24.878−16.596 1.00 30.46 C ATOM 673 C HIS A 88 0.029 25.581 −13.850 1.00 27.84C ATOM 674 O HIS A 88 −1.069 25.955 −14.267 1.00 26.38 O ATOM 675 N SERA 89 1.077 26.386 −13.719 1.00 26.97 N ATOM 676 CA SER A 89 1.039 27.791−14.093 1.00 24.52 C ATOM 677 CB SER A 89 1.502 28.665 −12.932 1.0025.63 C ATOM 678 OG SER A 89 1.724 29.994 −13.371 1.00 25.88 O ATOM 679C SER A 89 1.957 28.039 −15.276 1.00 23.29 C ATOM 680 O SER A 89 3.12027.638 −15.260 1.00 22.35 O ATOM 681 N ASN A 90 1.449 28.721 −16.2951.00 21.58 N ATOM 682 CA ASN A 90 2.274 29.010 −17.462 1.00 21.62 C ATOM683 CB ASN A 90 1.414 29.220 −18.716 1.00 22.59 C ATOM 684 CG ASN A 900.716 30.562 −18.737 1.00 23.18 C ATOM 685 OD1 ASN A 90 0.464 31.166−17.695 1.00 23.09 O ATOM 686 ND2 ASN A 90 0.394 31.037 −19.935 1.0021.28 N ATOM 687 C ASN A 90 3.291 30.146 −17.262 1.00 20.61 C ATOM 688 OASN A 90 4.123 30.393 −18.131 1.00 19.35 O ATOM 689 N LYS A 91 3.22430.830 −16.120 1.00 20.18 N ATOM 690 CA LYS A 91 4.184 31.890 −15.7921.00 20.62 C ATOM 691 CB LYS A 91 3.508 33.265 −15.768 1.00 21.83 C ATOM692 CG LYS A 91 2.995 33.739 −17.132 1.00 22.74 C ATOM 693 CD LYS A 913.939 34.731 −17.811 1.00 25.51 C ATOM 694 CE LYS A 91 3.500 35.008−19.248 1.00 28.49 C ATOM 695 NZ LYS A 91 3.757 36.414 −19.675 1.0030.57 N ATOM 696 C LYS A 91 4.839 31.559 −14.447 1.00 21.15 C ATOM 697 OLYS A 91 4.158 31.154 −13.506 1.00 19.94 O ATOM 698 N SER A 92 6.15531.734 −14.359 1.00 20.82 N ATOM 699 CA SER A 92 6.918 31.308 −13.1791.00 22.01 C ATOM 700 CB SER A 92 8.420 31.449 −13.440 1.00 21.23 C ATOM701 OG SER A 92 8.777 32.804 −13.657 1.00 21.35 O ATOM 702 C SER A 926.559 32.018 −11.877 1.00 22.28 C ATOM 703 O SER A 92 6.937 31.583−10.792 1.00 22.45 O ATOM 704 N GLN A 93 5.807 33.098 −12.006 1.00 22.36N ATOM 705 CA GLN A 93 5.441 33.975 −10.906 1.00 23.73 C ATOM 706 CB GLNA 93 4.982 35.266 −11.557 1.00 25.56 C ATOM 707 CG GLN A 93 5.433 35.222−13.015 1.00 29.43 C ATOM 708 CD GLN A 93 5.048 36.419 −13.837 1.0033.72 C ATOM 709 OE1 GLN A 93 3.981 36.998 −13.647 1.00 35.59 O ATOM 710NE2 GLN A 93 5.911 36.785 −14.777 1.00 37.81 N ATOM 711 C GLN A 93 4.35733.374 −10.013 1.00 22.06 C ATOM 712 O GLN A 93 4.135 33.830 −8.888 1.0019.00 O ATOM 713 N TYR A 94 3.687 32.347 −10.528 1.00 22.03 N ATOM 714CA TYR A 94 2.668 31.631 −9.771 1.00 22.48 C ATOM 715 CB TYR A 94 1.26931.861 −10.345 1.00 22.29 C ATOM 716 CG TYR A 94 0.912 33.322 −10.3421.00 25.27 C ATOM 717 CD1 TYR A 94 1.057 34.087 −11.487 1.00 23.39 CATOM 718 CE1 TYR A 94 0.757 35.429 −11.488 1.00 26.39 C ATOM 719 CZ TYRA 94 0.316 36.029 −10.328 1.00 26.49 C ATOM 720 OH TYR A 94 0.016 37.370−10.332 1.00 29.72 O ATOM 721 CE2 TYR A 94 0.174 35.293 −9.171 1.0025.15 C ATOM 722 CD2 TYR A 94 0.478 33.948 −9.181 1.00 22.81 C ATOM 723C TYR A 94 3.016 30.156 −9.695 1.00 22.66 C ATOM 724 O TYR A 94 3.69829.612 −10.566 1.00 21.05 O ATOM 725 N GLU A 95 2.515 29.526 −8.643 1.0023.58 N ATOM 726 CA GLU A 95 2.858 28.172 −8.272 1.00 25.71 C ATOM 727CB GLU A 95 3.845 28.302 −7.114 1.00 27.10 C ATOM 728 CG GLU A 95 4.92627.255 −6.935 1.00 34.72 C ATOM 729 CD GLU A 95 5.492 27.329 −5.528 1.0044.27 C ATOM 730 OE1 GLU A 95 6.286 28.253 −5.241 1.00 46.74 O ATOM 731OE2 GLU A 95 5.106 26.486 −4.693 1.00 47.77 O ATOM 732 C GLU A 95 1.58327.543 −7.732 1.00 25.85 C ATOM 733 O GLU A 95 0.818 28.213 −7.041 1.0024.94 O ATOM 734 N ILE A 96 1.338 26.270 −8.029 1.00 23.73 N ATOM 735 CAILE A 96 0.178 25.591 −7.452 1.00 24.08 C ATOM 736 CB ILE A 96 −0.74125.000 −8.536 1.00 24.17 C ATOM 737 CG1 ILE A 96 −1.363 26.102 −9.3881.00 21.98 C ATOM 738 CD1 ILE A 96 −0.539 26.449 −10.594 1.00 21.05 CATOM 739 CG2 ILE A 96 −1.821 24.131 −7.896 1.00 22.07 C ATOM 740 C ILE A96 0.566 24.454 −6.520 1.00 24.93 C ATOM 741 O ILE A 96 1.389 23.611−6.871 1.00 24.71 O ATOM 742 N ILE A 97 −0.033 24.425 −5.334 1.00 24.49N ATOM 743 CA ILE A 97 0.145 23.284 −4.450 1.00 24.95 C ATOM 744 CB ILEA 97 0.240 23.708 −2.985 1.00 25.07 C ATOM 745 CG1 ILE A 97 1.347 24.744−2.808 1.00 26.12 C ATOM 746 CD1 ILE A 97 1.587 25.126 −1.372 1.00 28.90C ATOM 747 CG2 ILE A 97 0.505 22.493 −2.105 1.00 25.26 C ATOM 748 C ILEA 97 −1.035 22.344 −4.655 1.00 25.56 C ATOM 749 O ILE A 97 −2.193 22.766−4.624 1.00 25.85 O ATOM 750 N THR A 98 −0.733 21.073 −4.869 1.00 26.48N ATOM 751 CA THR A 98 −1.760 20.078 −5.131 1.00 26.12 C ATOM 752 CB THRA 98 −1.566 19.495 −6.538 1.00 27.13 C ATOM 753 OG1 THR A 98 −1.73720.525 −7.517 1.00 30.30 O ATOM 754 CG2 THR A 98 −2.671 18.500 −6.8601.00 27.06 C ATOM 755 C THR A 98 −1.736 18.928 −4.143 1.00 26.04 C ATOM756 O THR A 98 −0.762 18.181 −4.076 1.00 25.26 O ATOM 757 N GLN A 99−2.804 18.784 −3.371 1.00 25.89 N ATOM 758 CA GLN A 99 −2.936 17.613−2.520 1.00 27.34 C ATOM 759 CB GLN A 99 −2.498 17.874 −1.083 1.00 29.07C ATOM 760 CG GLN A 99 −3.374 18.799 −0.293 1.00 35.42 C ATOM 761 CD GLNA 99 −2.568 19.548 0.732 1.00 44.28 C ATOM 762 OE1 GLN A 99 −1.35019.383 0.800 1.00 47.59 O ATOM 763 NE2 GLN A 99 −3.230 20.380 1.524 1.0050.15 N ATOM 764 C GLN A 99 −4.353 17.081 −2.599 1.00 27.53 C ATOM 765 OGLN A 99 −5.314 17.740 −2.198 1.00 26.16 O ATOM 766 N GLY A 100 −4.46715.882 −3.150 1.00 27.07 N ATOM 767 CA GLY A 100 −5.756 15.267 −3.3511.00 29.12 C ATOM 768 C GLY A 100 −5.809 13.841 −2.870 1.00 29.52 C ATOM769 O GLY A 100 −4.792 13.153 −2.766 1.00 28.98 O ATOM 770 N GLY A 101−7.028 13.393 −2.609 1.00 30.03 N ATOM 771 CA GLY A 101 −7.257 12.086−2.046 1.00 30.82 C ATOM 772 C GLY A 101 −7.296 10.884 −2.960 1.00 32.07C ATOM 773 O GLY A 101 −7.501 10.960 −4.174 1.00 29.69 O ATOM 774 N SERA 102 −7.087 9.785 −2.276 1.00 34.25 N ATOM 775 CA SER A 102 −7.1258.468 −2.825 1.00 36.02 C ATOM 776 CB SER A 102 −6.193 7.556 −2.040 1.0036.55 C ATOM 777 OG SER A 102 −6.010 8.024 −0.722 1.00 39.12 O ATOM 778C SER A 102 −8.563 8.000 −2.713 1.00 35.90 C ATOM 779 O SER A 102 −9.3438.647 −2.018 1.00 35.23 O ATOM 780 N GLY A 103 −8.986 6.930 −3.365 1.0037.33 N ATOM 781 CA GLY A 103 −10.352 6.532 −3.114 1.00 37.60 C ATOM 782C GLY A 103 −11.382 6.699 −4.242 1.00 38.60 C ATOM 783 O GLY A 103−11.059 6.692 −5.432 1.00 38.63 O ATOM 784 N ASN A 104 −12.630 6.838−3.815 1.00 38.81 N ATOM 785 CA ASN A 104 −13.793 6.857 −4.707 1.0038.65 C ATOM 786 CB ASN A 104 −14.664 5.667 −4.347 1.00 39.50 C ATOM 787CG ASN A 104 −15.731 5.451 −5.372 1.00 42.05 C ATOM 788 OD1 ASN A 104−15.831 6.200 −6.336 1.00 51.15 O ATOM 789 ND2 ASN A 104 −16.528 4.419−5.186 1.00 45.02 N ATOM 790 C ASN A 104 −14.564 8.165 −4.687 1.00 37.26C ATOM 791 O ASN A 104 −14.755 8.806 −5.733 1.00 39.29 O ATOM 792 N GLNA 105 −15.023 8.554 −3.500 1.00 34.32 N ATOM 793 CA GLN A 105 −15.5899.893 −3.351 1.00 31.80 C ATOM 794 CB GLN A 105 −16.652 10.016 −2.2511.00 31.93 C ATOM 795 CG GLN A 105 −18.006 9.358 −2.498 1.00 27.94 CATOM 796 CD GLN A 105 −18.915 9.453 −1.266 1.00 25.21 C ATOM 797 OE1 GLNA 105 −18.712 8.763 −0.285 1.00 22.71 O ATOM 798 NE2 GLN A 105 −19.97810.255 −1.128 1.00 24.00 N ATOM 799 C GLN A 105 −14.336 10.676 −2.9331.00 31.46 C ATOM 800 O GLN A 105 −14.035 10.784 −1.743 1.00 31.00 OATOM 801 N SER A 106 −13.594 11.236 −3.894 1.00 29.69 N ATOM 802 CA SERA 106 −12.336 11.860 −3.487 1.00 28.53 C ATOM 803 CB SER A 106 −11.20311.262 −4.302 1.00 27.95 C ATOM 804 OG SER A 106 −11.201 11.780 −5.6161.00 30.97 O ATOM 805 C SER A 106 −12.273 13.375 −3.493 1.00 27.39 CATOM 806 O SER A 106 −13.071 14.071 −4.127 1.00 28.69 O ATOM 807 N HIS A107 −11.297 13.867 −2.743 1.00 25.64 N ATOM 808 CA HIS A 107 −11.13315.290 −2.536 1.00 25.50 C ATOM 809 CB HIS A 107 −11.417 15.601 −1.0671.00 25.52 C ATOM 810 CG HIS A 107 −12.176 16.870 −0.848 1.00 29.94 CATOM 811 ND1 HIS A 107 −13.478 16.889 −0.395 1.00 30.47 N ATOM 812 CE1HIS A 107 −13.887 18.140 −0.289 1.00 36.42 C ATOM 813 NE2 HIS A 107−12.897 18.934 −0.654 1.00 39.36 N ATOM 814 CD2 HIS A 107 −11.814 18.165−1.006 1.00 35.04 C ATOM 815 C HIS A 107 −9.729 15.758 −2.889 1.00 23.45C ATOM 816 O HIS A 107 −8.741 15.124 −2.522 1.00 23.20 O ATOM 817 N VALA 108 −9.650 16.871 −3.608 1.00 22.60 N ATOM 818 CA VAL A 108 −8.36617.466 −3.947 1.00 23.16 C ATOM 819 CB VAL A 108 −7.963 17.205 −5.4191.00 23.67 C ATOM 820 CG1 VAL A 108 −9.056 17.657 −6.374 1.00 19.49 CATOM 821 CG2 VAL A 108 −6.634 17.885 −5.747 1.00 24.42 C ATOM 822 C VALA 108 −8.397 18.962 −3.673 1.00 23.19 C ATOM 823 O VAL A 108 −9.39219.636 −3.947 1.00 22.31 O ATOM 824 N THR A 109 −7.312 19.471 −3.1011.00 23.42 N ATOM 825 CA THR A 109 −7.188 20.898 −2.854 1.00 23.93 CATOM 826 CB THR A 109 −6.990 21.183 −1.353 1.00 24.81 C ATOM 827 OG1 THRA 109 −8.164 20.791 −0.633 1.00 25.76 O ATOM 828 CG2 THR A 109 −6.91722.679 −1.101 1.00 24.77 C ATOM 829 C THR A 109 −6.042 21.496 −3.6661.00 23.53 C ATOM 830 O THR A 109 −4.908 21.020 −3.616 1.00 23.96 O ATOM831 N TYR A 110 −6.363 22.537 −4.430 1.00 23.10 N ATOM 832 CA TYR A 110−5.374 23.280 −5.199 1.00 22.02 C ATOM 833 CB TYR A 110 −5.856 23.506−6.627 1.00 22.66 C ATOM 834 CG TYR A 110 −6.201 22.260 −7.401 1.0025.02 C ATOM 835 CD1 TYR A 110 −7.524 21.882 −7.583 1.00 21.75 C ATOM836 CE1 TYR A 110 −7.849 20.755 −8.303 1.00 21.69 C ATOM 837 CZ TYR A110 −6.847 19.990 −8.858 1.00 22.49 C ATOM 838 OH TYR A 110 −7.17118.863 −9.576 1.00 23.78 O ATOM 839 CE2 TYR A 110 −5.525 20.348 −8.6981.00 22.23 C ATOM 840 CD2 TYR A 110 −5.209 21.478 −7.976 1.00 22.38 CATOM 841 C TYR A 110 −5.174 24.644 −4.551 1.00 21.65 C ATOM 842 O TYR A110 −6.114 25.430 −4.450 1.00 19.62 O ATOM 843 N THR A 111 −3.956 24.928−4.111 1.00 20.52 N ATOM 844 CA THR A 111 −3.670 26.218 −3.504 1.0020.25 C ATOM 845 CB THR A 111 −2.942 26.036 −2.152 1.00 22.42 C ATOM 846OG1 THR A 111 −3.807 25.340 −1.243 1.00 21.55 O ATOM 847 CG2 THR A 111−2.730 27.373 −1.458 1.00 23.51 C ATOM 848 C THR A 111 −2.864 27.019−4.520 1.00 20.64 C ATOM 849 O THR A 111 −1.736 26.658 −4.858 1.00 19.47O ATOM 850 N ILE A 112 −3.479 28.069 −5.057 1.00 18.73 N ATOM 851 CA ILEA 112 −2.831 28.887 −6.074 1.00 20.13 C ATOM 852 CB ILE A 112 −3.84129.484 −7.071 1.00 21.04 C ATOM 853 CG1 ILE A 112 −5.043 28.556 −7.2641.00 18.50 C ATOM 854 CD1 ILE A 112 −4.681 27.150 −7.686 1.00 20.39 CATOM 855 CG2 ILE A 112 −3.152 29.784 −8.399 1.00 22.39 C ATOM 856 C ILEA 112 −2.095 30.004 −5.385 1.00 21.15 C ATOM 857 O ILE A 112 −2.70530.885 −4.783 1.00 21.07 O ATOM 858 N GLN A 113 −0.777 29.983 −5.4851.00 18.89 N ATOM 859 CA GLN A 113 −0.012 30.967 −4.760 1.00 21.47 CATOM 860 CB GLN A 113 0.627 30.316 −3.539 1.00 20.84 C ATOM 861 CG GLN A113 1.638 29.256 −3.907 1.00 28.15 C ATOM 862 CD GLN A 113 2.153 28.503−2.707 1.00 28.39 C ATOM 863 OE1 GLN A 113 1.617 28.634 −1.607 1.0030.14 O ATOM 864 NE2 GLN A 113 3.196 27.709 −2.911 1.00 25.13 N ATOM 865C GLN A 113 1.065 31.673 −5.545 1.00 21.44 C ATOM 866 O GLN A 113 1.39531.341 −6.686 1.00 20.90 O ATOM 867 N THR A 114 1.593 32.676 −4.866 1.0021.33 N ATOM 868 CA THR A 114 2.691 33.487 −5.311 1.00 24.29 C ATOM 869CB THR A 114 2.755 34.811 −4.549 1.00 23.47 C ATOM 870 OG1 THR A 1142.945 34.553 −3.153 1.00 30.54 O ATOM 871 CG2 THR A 114 1.464 35.594−4.733 1.00 27.33 C ATOM 872 C THR A 114 3.948 32.645 −5.153 1.00 22.90C ATOM 873 O THR A 114 4.111 31.958 −4.145 1.00 23.43 O ATOM 874 N THR A115 4.829 32.665 −6.146 1.00 21.93 N ATOM 875 CA THR A 115 6.031 31.847−6.056 1.00 23.25 C ATOM 876 CB THR A 115 6.693 31.672 −7.423 1.00 24.35C ATOM 877 OG1 THR A 115 5.779 31.034 −8.322 1.00 27.30 O ATOM 878 CG2THR A 115 7.944 30.815 −7.303 1.00 22.44 C ATOM 879 C THR A 115 7.02532.470 −5.084 1.00 22.76 C ATOM 880 O THR A 115 7.145 33.692 −4.998 1.0022.71 O ATOM 881 N SER A 116 7.710 31.626 −4.325 1.00 21.39 N ATOM 882CA SER A 116 8.687 32.111 −3.365 1.00 23.10 C ATOM 883 CB SER A 1168.579 31.354 −2.044 1.00 22.58 C ATOM 884 OG SER A 116 7.421 31.740−1.329 1.00 20.00 O ATOM 885 C SER A 116 10.076 31.951 −3.938 1.00 23.62C ATOM 886 O SER A 116 10.294 31.154 −4.850 1.00 22.16 O ATOM 887 N SERA 117 11.022 32.713 −3.411 1.00 24.19 N ATOM 888 CA SER A 117 12.38032.605 −3.901 1.00 25.55 C ATOM 889 CB SER A 117 13.042 33.978 −4.0321.00 25.43 C ATOM 890 OG SER A 117 13.961 34.208 −2.981 1.00 27.71 OATOM 891 C SER A 117 13.174 31.693 −2.982 1.00 25.15 C ATOM 892 O SER A117 13.219 31.887 −1.767 1.00 24.64 O ATOM 893 N ARG A 118 13.793 30.689−3.583 1.00 24.99 N ATOM 894 CA ARG A 118 14.577 29.708 −2.857 1.0025.50 C ATOM 895 CB ARG A 118 14.675 28.442 −3.707 1.00 26.03 C ATOM 896CG ARG A 118 15.157 27.214 −2.979 1.00 27.66 C ATOM 897 CD ARG A 11814.905 25.928 −3.743 1.00 33.83 C ATOM 898 NE ARG A 118 13.724 25.209−3.270 1.00 32.39 N ATOM 899 CZ ARG A 118 12.605 25.059 −3.968 1.0039.89 C ATOM 900 NH1 ARG A 118 12.498 25.587 −5.180 1.00 43.31 N ATOM901 NH2 ARG A 118 11.590 24.376 −3.455 1.00 37.18 N ATOM 902 C ARG A 11815.978 30.208 −2.486 1.00 25.23 C ATOM 903 O ARG A 118 16.667 30.824−3.299 1.00 25.07 O ATOM 904 N TYR A 119 16.371 29.955 −1.240 1.00 25.20N ATOM 905 CA TYR A 119 17.715 30.232 −0.739 1.00 25.74 C ATOM 906 CBTYR A 119 17.638 30.485 0.775 1.00 24.93 C ATOM 907 CG TYR A 119 18.88130.905 1.579 1.00 26.32 C ATOM 908 CD1 TYR A 119 19.124 32.246 1.8811.00 26.48 C ATOM 909 CE1 TYR A 119 20.218 32.631 2.663 1.00 26.71 CATOM 910 CZ TYR A 119 21.065 31.671 3.180 1.00 28.22 C ATOM 911 OH TYR A119 22.142 32.049 3.952 1.00 30.00 O ATOM 912 CE2 TYR A 119 20.83130.334 2.923 1.00 27.78 C ATOM 913 CD2 TYR A 119 19.735 29.956 2.1401.00 25.89 C ATOM 914 C TYR A 119 18.433 28.925 −1.039 1.00 24.89 C ATOM915 O TYR A 119 19.657 28.851 −1.065 1.00 28.03 O ATOM 916 O HOH W 1−14.559 14.990 0.123 0.50 27.84 O ATOM 917 O HOH W 2 −3.227 22.536−1.713 1.00 24.73 O ATOM 918 O HOH W 3 2.121 20.187 −4.665 1.00 38.18 OATOM 919 O HOH W 4 −11.411 30.080 −22.842 1.00 30.93 O ATOM 920 O HOH W5 −13.591 32.301 −21.281 1.00 42.14 O ATOM 921 O HOH W 6 −2.445 23.591−19.928 1.00 36.28 O ATOM 922 O HOH W 7 −1.738 26.722 −17.284 1.00 43.23O ATOM 923 O HOH W 8 −18.754 29.155 −21.318 1.00 45.28 O ATOM 924 O HOHW 9 1.119 39.224 −7.410 1.00 44.97 O ATOM 925 O HOH W 10 0.534 33.622−2.189 1.00 32.51 O ATOM 927 O HOH W 11 7.824 25.984 −3.897 1.00 43.51 OATOM 928 O HOH W 12 19.203 30.879 −5.626 1.00 44.10 O ATOM 929 O HOH W13 −22.689 2.969 −19.990 1.00 42.82 O ATOM 930 O HOH W 14 −7.161 −0.951−10.204 1.00 38.86 O ATOM 932 O HOH W 15 −3.700 36.274 −1.163 1.00 36.83O ATOM 933 O HOH W 16 −23.572 24.381 −10.521 1.00 37.83 O ATOM 935 O HOHW 17 −20.762 13.499 −1.986 1.00 44.73 O ATOM 936 O HOH W 18 18.05832.923 −5.866 1.00 38.33 O ATOM 937 O HOH W 19 3.068 37.314 −2.600 1.0039.55 O ATOM 938 O HOH W 20 −5.029 22.918 2.276 1.00 36.67 O ATOM 939 OHOH W 21 7.686 32.276 −17.226 1.00 36.19 O ATOM 940 O HOH W 22 −10.31039.058 −5.931 1.00 39.25 O ATOM 941 O HOH W 23 5.409 29.664 −3.323 1.0031.60 O ATOM 942 O HOH W 24 −3.061 2.513 −9.205 1.00 42.76 O ATOM 943 OHOH W 25 −17.878 8.534 −16.806 1.00 38.00 O ATOM 944 O HOH W 26 −14.46632.442 −24.824 1.00 42.24 O ATOM 945 O HOH W 27 −17.952 26.616 −2.0861.00 38.39 O ATOM 946 O HOH W 28 −7.866 41.166 −9.335 1.00 37.60 O ATOM947 O HOH W 29 −5.147 25.608 0.944 1.00 26.31 O ATOM 948 O HOH W 303.398 25.915 −12.359 1.00 42.41 O ATOM 950 O HOH W 31 3.685 17.090−11.869 1.00 43.77 O ATOM 952 O HOH W 32 −6.148 2.482 −6.226 1.00 45.41O ATOM 953 O HOH W 33 −1.142 26.694 −20.196 1.00 46.61 O ATOM 954 O HOHW 34 −5.285 32.131 −22.226 1.00 43.31 O ATOM 955 O HOH W 35 −11.27341.495 −9.609 1.00 35.70 O ATOM 956 O HOH W 36 −5.031 44.491 −12.0011.00 36.83 O ATOM 957 O HOH W 37 −7.969 25.331 1.388 1.00 44.78 O ATOM958 O HOH W 38 3.127 24.741 −9.731 1.00 39.57 O ATOM 959 O HOH W 3920.331 30.525 −1.815 1.00 38.12 O ATOM 960 O HOH W 40 −15.691 35.326−5.763 1.00 45.12 O ATOM 961 O HOH W 41 9.232 23.005 −2.062 1.00 36.89 OATOM 962 O HOH W 42 5.969 28.000 −14.974 1.00 41.48 O ATOM 963 O HOH W43 3.257 41.375 −6.139 1.00 42.08 O ATOM 964 O HOH W 44 5.399 27.827−11.600 1.00 45.55 O ATOM 965 O HOH W 45 0.775 21.354 −8.403 1.00 37.66O ATOM 967 O HOH W 46 −21.248 26.695 −8.253 1.00 40.43 O ATOM 968 O HOHW 47 −8.574 41.852 −5.684 1.00 40.41 O ATOM 969 O HOH W 48 −24.65923.245 −6.310 1.00 40.25 O ATOM 970 O HOH W 49 −17.132 6.949 −0.155 1.0039.44 O ATOM 972 O HOH W 50 −3.415 28.071 −24.096 1.00 44.88 O ATOM 973O HOH W 51 −20.139 22.500 −2.785 1.00 44.66 O ATOM 974 O HOH W 52 −0.98637.218 −2.381 1.00 44.32 O ATOM 976 O HOH W 53 −22.382 3.698 −22.5641.00 42.32 O ATOM 979 O HOH W 54 8.478 38.108 −4.765 1.00 40.63 O ATOM982 O HOH W 55 −19.946 7.127 −4.521 1.00 43.21 O ATOM 984 O HOH W 566.496 39.021 −16.893 1.00 44.40 O ATOM 985 O HOH W 57 −17.455 18.329−1.316 1.00 46.16 O ATOM 987 O HOH W 58 −12.524 33.301 −24.061 1.0040.56 O ATOM 988 O HOH W 59 6.414 37.040 −4.985 1.00 40.47 O ATOM 989 OHOH W 60 −9.023 34.535 −20.227 1.00 44.58 O ATOM 990 O HOH W 61 −27.46422.652 −5.829 1.00 38.44 O ATOM 991 O HOH W 62 −26.909 14.965 −7.8931.00 41.51 O ATOM 992 O HOH W 63 −24.626 6.229 −11.331 1.00 45.20 O ATOM993 O HOH W 64 −17.082 24.917 −19.027 1.00 42.52 O ATOM 994 O HOH W 65−3.009 28.610 −26.134 1.00 41.64 O ATOM 995 O HOH W 66 −15.913 17.226−27.554 1.00 41.28 O ATOM 996 O HOH W 67 3.707 23.828 −13.461 1.00 41.38O

APPENDIX 2 PLOTSIMILARITY of:/ul/home/burmeist/projects/binaries/14kD/14kDrops.msf(*) from: 1 to: 133Window: 1 Scoring matrix: GenRunData: blosum62.cmp March 28, 2002 15:28Accessible: 0 < 30%, 1 >= 30%, 2 >= 50%, 3 >= 80% Out-facing: 0 not, 1sort of, 2 obvious B-factor: <=20, Obs B- sub- factor B-factorstitutions 0-5 residue (-* Similarity Out- cool to avg from PositionConsensus Cry34Ab1 invariant) Sub-stitutions Value Acc facing hot PDBfile 1 M M — ML(IV)(FQ) 5.0 — — — 2 S S — S(TNA)(DEGKQ) 4.0 — — — 3 A A— AGS|T(VC)EQP 4.0 2 2 2 31.14 4 R R — RKQN|(HE)G(SA) 5.0 0 1 3 34.86 5E E — EQDKR(HNS) 5.0 0 2 5 36.48 6 V V — VILM|(AT) 4.0 0 0 1 24.20 7 H H— HYN 8.0 0 2 1 21.39 8 I I — IVL 4.0 0 0 1 22.99 9 E D DEQNS|KH 2.8 0 21 24.03 10 I V VILM|ATPCY 3.4 0 0 1 23.49 11 N N I NS(HD)|T(QRE) 2.4 0 21 21.80 12 N N — N(DSH)(EGKQT) 6.0 0 0 1 21.90 13 K K H KR(QE)N|SH 2.8 12 1 21.76 14 T T — TS(VN) 5.0 0 0 1 22.87 15 G G R GN|(AS)R 1.9 0 2 126.60 16 H H — HYN 8.0 0 2 2 29.89 17 T T — TS(ANV) 5.0 0 2 1 26.99 18 LL — L(IM)VF 4.0 0 0 1 24.62 19 Q Q — QE(RK)(SMND) 5.0 1 2 1 23.34 20 L LM LMIV|P(AT)C 3.3 0 2 2 26.00 21 E E D EDQKNS|HR 3.9 0 2 3 31.91 22 D DK DENQ 3.3 2 2 2 27.82 23 K K R KRQEN|S 3.8 1 2 2 28.57 24 T T — TS(VNA)5.0 0 1 1 24.63 25 K K R KRQEN|S 3.8 1 2 2 29.78 26 L L — L(MI)VF 4.0 00 1 26.83 27 D D S, A, T DESNT|QAGP 1.2 1 2 2 30.83 28 G G H S GSN|A 2.90 1 1 26.65 29 G G — G(NSA) 6.0 0 1 1 25.27 30 R R E RKE(NH) 3.1 1 2 229.56 31 W W — WY 11.0 0 0 2 26.87 32 R R I RKQ|(NHE) 1.9 1 2 2 27.54 33T T I TSV|ANI 2.6 0 1 1 27.09 34 S S T ST(NA)|(DEKQ)G 2.9 0 2 1 26.59 35P P — P 7.0 0 0 1 26.11 36 T T V TSVA|(N) 3.0 0 2 2 26.81 37 N N —N(DHS)(EGKQRT) 6.0 1 2 1 22.77 38 V V — VI(LM)(AT) 4.0 0 0 1 23.17 39 AA P AS|T(VG)(CP) 2.1 0 2 1 23.75 40 N N R NRHS 2.9 0 1 1 26.09 41 D D NDENSQ 4.1 1 2 1 24.68 42 Q Q S, T QTESKRN|DMAP 1.2 0 2 1 21.44 43 I I SIVLM|F(AT)C 1.7 2 2 2 25.76 44 K K D KREQNS|D 2.7 0 2 3 27.98 45 T T LTSV|ANL 2.6 0 2 1 23.97 46 F F — FYWILK 6.0 0 0 1 25.47 47 V V QVIML|(AT)Y 1.7 0 2 T45, 1 24.30 E49, R31, E4, H7 cover 48 A A — ASCGTV4.0 0 0 2 29.93 49 E E G EDQK(SN)|(RH) 2.3 0 2 3 33.24 50 S S —S(TNA)(DEGKQ) 4.0 0 0 2 31.97 51 H N D HYN 3.4 1 2 4 40.32 52 G G —G(ANS) 6.0 0 2 2 31.85 53 F F V PYWI(LM) 3.2 2 2 1 24.44 54 M M LMLIVF(Q) 3.8 1 2 2 27.92 55 T T — TS(VNA) 5.0 0 2 1 24.25 56 G G —G(ANS) 6.0 0 1 1 26.79 57 V T I TVI|LAMSC 1.9 0 0 2 28.35 58 E E —EDQK(HNRS) 5.0 0 2 2 27.64 59 G G — G(ANS) 6.0 0 0 1 24.40 60 I T HTIVL|M . . . 0.5 0 2 1 24.59 61 I I — IVLMF 4.0 0 0 1 23.37 62 Y Y IYF(WH) 3.8 0 2 1 25.43 63 Y Y F YFWH 4.9 0 0 2 24.79 64 S S T STA|. . .2.9 0 2 1 25.69 65 I I V IVLM 3.5 0 0 2 33.11 66 N N — N(DHS)(EKQRT) 6.00 2 5 44.24 67 G G — G(ANS) 6.0 1 2 2 37.39 68 E E D EDQKS|HR 3.6 0 2 234.17 69 A A I 2.1 0 0 1 24.99 70 E E — 5.0 0 2 2 26.75 71 I I — 4.0 0 02 25.87 72 S S T, P 2.3 0 2 1 23.17 73 L L — 4.0 0 0 1 21.67 74 H Y 4.50 2 2 24.32 75 F F — 6.0 0 0 1 22.92 76 D D — 6.0 0 2 side- 2 20.49chain coverage 77 N N — 6.0 0 0 0 18.23 78 P P — 7.0 0 2 0 20.59 79 Y F6.1 1 2 1 22.67 80 A A I, S 0.5 1 2 2 28.72 81 G G — 6.0 1 2 2 29.78 82S S — 4.0 1 2 2 26.29 83 N N — 6.0 0 1 x- 2 25.83 domain h-bonds 84 K K— 5.0 1 2 3 29.80 85 Y Y C 2.7 0 0 2 25.28 86 D D S 3.6 1 2 2 25.96 87 GG — 6.0 0 0 1 26.03 88 H H R, S, D 0.5 1 2 2 28.77 89 S S — 4.0 0 0 124.77 90 N N D, S 2.2 1 2 1 21.66 91 K K D 2.7 1 2 2 23.44 92 P S D, N1.5 1 2 0 21.69 93 Q Q E, D 2.4 1 2 2 27.69 94 Y Y — 7.0 0 0 2 24.14 95E E K 3.4 0 2 4 33.40 96 V I 3.8 0 0 1 23.34 97 I I T 2.1 0 2 2 25.77 98T T — 5.0 0 1 2 26.91 99 Q Q E 3.8 1 2 5 34.82 100 G G S, A 1.6 0 2 228.67 101 R — 0.1 — — — — 102 A — 0.1 — — — — 103 E — 0.1 — — — — 104 H— 0.2 — — — — 105 G G R 2.9 0 0 2 30.65 106 S S A 2.8 2 2 2 36.17 107 GG N 3.7 0 2 2 38.04 108 N N D 3.5 0 1 5 41.46 109 Q Q K, H 1.6 2 2 228.93 110 S S D 2.5 0 0 2 28.87 111 H H — 8.0 0 2 2 29.45 112 V V — 4.00 0 1 22.69 113 T T — 5.0 0 2 1 24.31 114 Y Y — 7.0 0 0 1 22.36 115 T T— 5.0 0 2 1 21.19 116 I I V 3.6 0 0 1 20.42 117 Q Q — 5.0 0 2 2 23.92118 T T R 2.7 1 2 2 24.75 119 V T N, A 0.1 0 2 1 23.53 120 S S I 1.7 1 21 22.14 121 S S L 1.0 2 2 1 25.44 122 R R — 5.0 2 2 3 31.00 123 Y Y L2.8 3 2 2 26.68 124 G G *, T −1.2 — 125 N H 1.8 — 126 N K 0.9 — 127 S SL 0.3 — 128 C −0.5 129 S 0.1 130 N 0.2 131 N 0.2 132 S 0.1 133 * 0.0

1. A truncated Cry34 protein comprising residues 1-114 of a wild-typeCry34 protein and having at least one deletion after residue
 114. 2. Thetruncated Cry34 protein of claim 1 comprising residues 1-118 of saidwild-type Cry34 protein.
 3. A method of inhibiting a rootworm, whereinsaid method comprises providing, for ingestion, to said rootworm theprotein according to claim 1.